Macrocyclic compounds and uses thereof

ABSTRACT

The present disclosure relates to certain macrocyclic compounds that inhibit SRC and MET, and/or CSF1R, pharmaceutical compositions containing such compounds, and methods of using such compounds to treat cancer.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority under 35 U.S.C. § 119(e) to U.S.Provisional Application Ser. No. 62/538,193 filed on Jul. 28, 2017, andU.S. Provisional Application Ser. No. 62/700,990 filed on Jul. 20, 2018,the entire disclosures of which are incorporated herein by reference.

TECHNICAL FIELD

The present disclosure relates to certain macrocyclic compounds thatinhibit SRC and MET, and/or CSF1R, pharmaceutical compositionscontaining such compounds, and methods of using such compounds to treatcancer.

BACKGROUND

Protein kinases are key regulators for cell growth, proliferation andsurvival. Genetic and epigenetic alterations accumulate in cancer cellsleading to abnormal activation of signal transduction pathways whichdrive malignant processes. (Manning, G. et al, The protein kinasecomplement of the human genome. Science 2002, 298, 1912-1934).Pharmacological inhibition of these signaling pathways presentspromising intervention opportunities for targeted cancer therapies.(Sawyers, C. Targeted cancer therapy. Nature 2004, 432, 294-297).

MET, also called hepatocyte growth factor receptor (HGFR), wasdiscovered in 1984 (Cooper, C. S., et al Molecular cloning of a newtransforming gene from a chemically transformed human cell line. Nature1984, 311, 29-33). Hepatocyte growth factor (HGF), also known as scatterfactor (SF), is the high-affinity natural ligand of MET (Bottaro D P etal. Identification of the hepatocyte growth factor receptor as the c-netproto-oncogene product. Science. 1991, 251 (4995), 802-804). The HGF/METsignaling pathway is implicated in invasive growth during embryodevelopment, postnatal organ regeneration, wound healing and tissueregeneration processes. However, the HGF/MET axis is frequently hijackedby cancer cells for tumorigenesis, invasive growth, and metastasis(Boccaccio, C.; Comoglio, P. M. Invasive growth: a MET-driven genericprogramme for cancer and stem cells. Nat. Rev. Cancer 2006, 6, 637-645).Deregulations of MET and/or HGF via activating mutations, geneamplifications, overexpression, and both autocrine or paracrine loopregulation influence cell growth, proliferation, angiogenesis, invasion,survival, and metastasis, leading to tumorigenesis and tumor progression(Ma, P C et al. Expression and mutational analysis of MET in human solidcancers. Genes Chromosomes Cancer 2008, 47, 1025-1037). Over-expressionof MET and/or HGF has been detected in a large variety of solid tumorssuch as liver, breast, pancreas, lung, kidney, bladder, ovary, brain,prostate, and many others, and is often associated with a metastaticphenotype and poor prognosis (Maulik, G., et al. Role of the hepatocytegrowth factor receptor, MET, in oncogenesis and potential fortherapeutic inhibition. Cytokine Growth Factor Rev. 2002, 13, 41-59).MET amplification has been reported in different human cancers includinggastroesophageal carcinomas, colorectal cancers, NSCLC,medulloblastomas, and glioblastomas (Smolen, G. A., et al. Amplificationof MET may identify a subset of cancers with extreme sensitivity to theselective tyrosine kinase inhibitor PHA-665752. Proc. Natl. Acad. Sci.U.S.A 2006, 103, 2316-2321). A diverse set of MET mutations in thetyrosine kinase domain, juxtamembrane, and extracellular domain of bothgermline and somatic mutations have been described in many solid tumors,including hereditary and sporadic human papillary renal carcinomas, lungcancer, ovarian cancer, childhood hepatocellular carcinomas, squamouscell carcinoma of the head and neck, and gastric cancer (Ghiso, E.;Giordano, S. Targeting MET: why, where and how? Curr. Opin. Pharmacol.2013, 13, 511-518). MET exon 14 deletion represents a novel as ofactionable oncogenic event with potential clinical impact andtherapeutic applications in patients affected by different cancer types(Pilotto S, MET exon 14 juxtamembrane splicing mutations: clinical andtherapeutical perspectives for cancer therapy. Ann Transl Med. 20175(1):2). Autocrine or paracrine stimulation is one mechanism foraberrant MET activation. The MET autocrine activation plays a causalrole in the development of malignant melanoma and acquisition of themetastatic phenotype (Otsuka, T., et al. MET autocrine activationinduces development of malignant melanoma and acquisition of themetastatic phenotype. Cancer Res. 1998, 58, 5157-5167). For glioblastoma(GBM), HGF autocrine expression correlated with MET phosphorylationlevels in HGF autocrine cell lines, and showed high sensitivity to METinhibition in vivo, while an HGF paracrine environment could enhanceglioblastoma growth in vivo but did not demonstrated sensitivity to METinhibition (Xie, Q., et al. Hepatocyte growth factor (HGF) autocrineactivation predicts sensitivity to MET inhibition in glioblastoma. Proc.Natl. Acad. Sci. U.S.A 2012, 109, 570-575). The aberrant expression ofHGF is a crucial element in AML pathogenesis that leads to autocrineactivation of MET in nearly half of the AML cell lines and clinicalsamples (Kentsis, A., et al. Autocrine activation of the MET receptortyrosine kinase in acute myeloid leukemia. Nat. Med. 2012, 18,1118-1122).

Upregulation of HGF/MET signaling has been frequently reported ascompensatory signaling to confer resistance for kinase targetedtherapies. MET amplification has been detected in 4%-20% of NSCLCpatients with the EGFR mutations who acquired resistance to gefitinib orerlotinib treatment (Sequist, L. V., et al. Analysis of tumor specimensat the time of acquired resistance to EGFR-TKI therapy in 155 patientswith EGFR-mutant lung cancers. Clin. Cancer Res. 2013, 19, 2240-2247).Upregulation of ligand HGF represents another mechanism of EGFR-TKIresistance. High HGF expression was discovered among clinical specimenswith acquired resistance that did not have a T790M mutation or METamplification as well as among cases that exhibited primary resistancedespite having EGFR-TKI sensitive activating EGFR gene mutations (Yano,S., et al. Hepatocyte growth factor induces gefitinib resistance of lungadenocarcinoma with epidermal growth factor receptor-activatingmutations. Cancer Res. 2008, 68, 9479-9487). Amplification of MET isassociated with acquired resistance to cetuximab or panitumumab inmetastatic colorectal cancer patients that do not develop KRAS mutationsduring anti-EGFR therapy (Bardelli, A., et al. Amplification of the METReceptor Drives Resistance to Anti-EGFR Therapies in Colorectal Cancer.Cancer Discov. 2013, 3, 658-673). Growth factor-driven resistance fromtumor microenvironment represents a potential common mechanism foranticancer kinase inhibitors. The upregulation of stromal HGF confersresistance to the BRAF inhibitor ramurafenib in BRAF-mutant melanomacells (Straussman, R., et al. Tumour micro-environment elicits innateresistance to RAF inhibitors through HGF secretion. Nature 2012, 487,500-504). It was reported that ligand-mediated activation of alternativereceptor tyrosine kinases was observed in cancer cells originallydependent on either MET, FGFR2, or FGFR3, and RTKs from the HER and EGFRfamilies as well as MET compensated for loss of each other (Harbinski,F., et al. Rescue screens with secreted proteins reveal compensatorypotential of receptor tyrosine kinases in driving cancer growth. CancerDiscov. 2012, 2, 948-959). Therefore, blocking adaptive cellularresponses that drive compensatory ligand expression is necessary forachieving optimal and sustained antitumor effects.

Oncogenic K-Ras mutation occurs frequently in cancers, includingpancreatic, gastric, and lung cancers. K-Ras mutant ancers are moredependent on K-Ras in anchorage-independent culture conditions than inmonolayer culture conditions. Enhanced Met expression and signaling isessential for anchorage-independent growth of K-Ras mutant cancer cellsand suggests that pharmacological inhibitors of MET could be effectivefor K-Ras mutant tumor patients (Fujita-Sato, S., et al. Enhanced METTranslation and Signaling Sustains K-Ras-Driven Proliferation underAnchorage-Independent Growth Conditions. Cancer Res. 2015, 75,2851-2862).

Cytoplasmic tyrosine kinases of the SRC family (SFKs) play importantroles in signal transduction induced by a large number of extracellularstimuli including growth factors and integrins (Parsons, S. J., et al.Src family kinases, key regulators of signal transduction. Oncogene,2004, 23, 7906-7909). Elevated expression of the nonreceptor tyrosinekinase SRC and/or increased SRC kinase activity has been reported in awide variety of human cancers, including breast, colon, lung, and headand neck cancers. Increased activation of SRC and STAT3 was reported tobe associated with many epithelial cancers and linked to the expressionof a number of growth factors such as vascular endothelial growth factorand HGF. SRC and STAT3 can act cooperatively as upstream regulators ofHGF expression, resulting in establishment of an HGF autocrine loop,signal amplification, and an invasive phenotype (Wojcik, E. J., et al. Anovel activating function of SRC and STAT3 on HGF transcription inmammary carcinoma cells. Oncogene. 2006, 25, 2773-84). Therefore,targeting SRC/STAT3-signalling pathway may be an effective fordisruption of autocrine HGF loops in cancers. EGFR inhibitors have goodresponse only in EGFR-mutant NSCLC patients. The wildtype EGFRactivation of invasive phenotypes rely largely on EGFR-SRC-MET signalingthrough HGF-independent pathway (Dulak A M, et al. HGF-independentpotentiation of EGFR action by MET. Oncogene. 2011, 30, 3625-3635). EGFRligands induce accumulation of activated MET, which begins at 8 h andcontinues for 48 h, leading to an increase in MET expression andphosphorylation of critical MET tyrosine residues without activation ofmitogen-activated protein kinase (MAPK) or AKT. This gene transcriptionrelated lateral signaling is associated with prolonged SRCphosphorylation, and the SRC pathway is involved with EGFR to METcommunication. Although EGFR is overexpressed in about 90% of head andneck squamous cell carcinoma (FINSCC), EGER inhibitors developed to-datehave been provided limited clinical efficacy. For example,ligand-independent activation of MET contributes specifically toerlotinib resistance in HNSCC with activated SRC, where MET activationis more dependent on SRC than on EGFR, providing an alternate survivalpathway (Stabile, L. P., et al. c-SRC activation mediates erlotinibresistance in head and neck cancer by stimulating MET. Clin Cancer Res.2012, 19, 1-13). Aberrant activation of SRC has been demonstrated innumerous epithelial tumors, including HNSCC. SRC inhibition resulted ina universal and profound reduction of invasion and migration of HNSCCcell lines, but produced cytotoxicity in some of HNSCC cell lines.Sustained MET activation mediates resistance to SRC inhibition. Thesynergistic cytotoxic effects of SRC and MET inhibition were observed inHNCC cell lines (Sen, B., et al. Distinct interactions between SRC andMET in mediating resistance to SRC inhibition in head and neck cancer.Clin Cancer Res. 2010, 17, 1-11).

It was reported that cetuximab-induced MET activation led to cetuximabresistance in Caco-2 colon cancer cells, and SRC activation promotedcetuximab resistance by interacting with MET via MET/SRC/EGFR complexformation (Song N, et al. Cetuximab-induced MET activation acts as anovel resistance mechanism in colon cancer cells. Int J Mol Sci. 2014,15, 5838-5851). SRC is a key downstream transducer of MET-driven tumorgrowth. Inhibition of SRC in Met-addicted gastric carcinoma cell linesenhanced the cell sensitivity to inhibition of MET that supports thetherapeutic potential of combinatorial treatment with MET and SRCinhibitors (Bertotti, A., et al. Inhibition of SRC impairs the growth ofMET-addicted gastric tumors. Clin Cancer Res. 2010, 16, 3933-3943).Although HGF/MET signaling is implicated in the development ofcolorectal cancer (CRC), inhibition of MET alone has been demonstratedto have limited efficacy. SRC activation was essential forligand-dependent and independent activation of MET. The combinedinhibition of MET and SRC enhanced the inhibition of cell proliferationand apoptosis in mutant and wild type RAS colon cancer cells (Song, N.,et al. Dual inhibition of MET and SRC kinase activity as a combinedtargeting strategy for colon cancer. Exp Ther Med etm.2017.4692).

CSF1R, also known as FMS, is a receptor for colony stimulating factor 1,a cytokine that controls the production, differentiation, and functionof macrophages. Non-resolving inflammation in the tumor microenvironmentis a hallmark of cancer and associated with M2-polarized macrophages.Tumor associated macrophages (TAMs) more closely resemble M2-polarizedmacrophages, and play important roles in promoting proliferation,invasion, and metastasis of cancer (Yang L, et al. Tumor-associatedmacrophages: from basic research to clinical application. J HematolOncol. 2017, 10, 58). The tumor-promoting function of TAMs is based ontheir capacity to secrete proangiogenic and growth factors, as well asto potently suppress T cell effector function by releasingimmunosuppressive cytokines and affecting their metabolism (Ries C H, etal. Targeting tumor-associated macrophages with anti-CSF1R antibodyreveals a strategy for cancer therapy. Cancer Cell. 2014, 25, 846-859).Although anti-PD-1 monoclonal antibodies (mAbs) targeting the immunecheckpoint have demonstrated benefits for the treatment of certaincancers, these drugs are not always effective. Recent studies indicatedthat the efficacy of anti-PD-1 mAbs was impacted by the uptake ofanti-PD-1 mAbs-bound PD-1+tumor-infiltrating CD8+ T cells byPD-1−tumor-associated macrophages. Combination therapies designed totarget tumor macrophages and anti-PD-1, may provide additional benefitby increasing immune checkpoint blockade drug delivery to CD8+ T cells,thereby enhancing activity of immunotherapy (Arlauckas S P, et al. Invivo imaging reveals a tumor-associated macrophage-mediated resistancepathway in anti-PD-1 therapy. Sci Transl Med. 2017, 9(389). pii:eaa13604). Survival of TAMs is mediated by signaling throughcolony-stimulating factor 1 receptor (CSF1R), and inhibition of CSF1Rsignaling reduces TAMs and increases CD8/CD4 T-cell ratio in patientswith advanced solid tumors. Therefore, targeting CSF1R signaling leadingto the modulation of TAMs is a promising therapeutic strategy in varioussolid tumors, as a single agent or in combination with standard of carechemotherapeutic agents and immunotherapies. Coexpression of CSFIR andCSH is most often detected in invasive tumors. Autocrine CSF-1Ractivation induced hyperproliferation and disruption of junctionalintegrity in acinar structures formed by human mammary epithelial cellsin three-dimensional culture through a SRC-dependent mechanism (Wrobel CN, et al. Autocrine CSF1R activation promotes SRC-dependent disruptionof mammary epithelial architecture. J Cell Biol. 2004, 165, 263-273).Inhibition of CSF-1R and SRC may prove to be a valuable strategy in thetreatment of invasive tumors. Tenosynovial giant cell tumor (TGCT) orpigmented villonodular synovitis (PVNS) is a clonal neoplasticproliferation arising from cells overexpressing CSF1 that recruitCSF1R-bearing polyclony macrophages and make up the bulk of the tumor.Inhibition of CSF1R using small molecule inhibitors can lead toimprovement in the affected joint (Ravi V, et al. Treatment oftenosynovial giant cell tumor and pigmented villonodular synovitis. CurrOpin Oncol. 2011, 23, 361-366).

In a summary, aberrant activation of HGF/MET pathway has frequently beenfound in human cancers via protein over-expression, mutation, geneamplification, and also paracrine or autocrine upregulation. Inaddition, the activation of HGF/MET signaling confers resistance tocancer therapies. SRC activation is implicated for ligand-dependent andindependent activation of MET. CSF1R plays an import role in regulationof tumor associated macrophage. Therefore, the polypharmacologicinhibition of MET/SRC/CSF1R has great potential for therapeuticinterventions in cancers. To-date, compounds that inhibit MET/SRC and/orCSF1R have been elusive. As such, there exists a significant unmet need.

SUMMARY

In one aspect, the disclosure relates to a compound of the Formula I

or a pharmaceutically acceptable salt thereof, wherein

X¹ and X² are independently CR⁶R⁷—, S, S(O), S(O)₂, O or N(R⁸);

R¹ is H, deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, C₃-C₁₀ aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein each hydrogenatom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl andC₆-C₁₀ aryl is independently optionally substituted by deuterium,halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂,—NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆alkyl, —NHC(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂,—NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆alkyl), —NHS(O)₂(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl), —NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆alkyl)S(O)NH₂, —N(C₁-C₆ alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl),—NHS(O)₂NH(C₁-C₆ alkyl), —NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆alkyl)₂, —N(C₁-C₆ alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂, —SC₁-C₆ alkyl, —S(O)C₁-C₆alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆ alkyl), —S(O)₂NH(C₁-C₆ alkyl),—S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆ alkyl)₂, —P(C₁-C₆alkyl)₂,—P(O)(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or 3- to 7-memberedheterocycloalkyl;

each R² and R³ is independently H, deuterium, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁸ or—C(O)NR⁸R⁹; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl and C₆-C₁₀ aryl is independentlyoptionally substituted by deuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl,—NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂,—N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆alkyl)₂, —P(O)(C₁-C₆alkyl)₂, C₃-C₆ cycloalkyl, or 3- to7-membered heterocycloalkyl; or R² and R³ taken together with the carbonatoms to which they are attached optionally form a C₅-C₇ cycloalkyl or a5- to 7-membered heterocycloalkyl; or R² and R⁴ taken together with theatoms to which they are attached optionally form a 5- to 7-memberedheterocycloalkyl;

R⁴ is H, C₁-C₆ alkyl or 3- to 7-membered heterocycloalkyl, wherein eachhydrogen atom in C₁-C₆ alkyl or 3- to 7-membered heterocycloalkyl isindependently optionally substituted by halogen, —OH, —CN, —OC₁-C₆alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —CO₂H, —C(O)OC₁-C₆alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂, C₃-C₆cycloalkyl, or monocyclic 5- to 7-membered heterocycloalkyl;

R⁵ is H or —NR⁶R⁷;

each R⁶, R⁷ and R⁸ are each independently selected from the groupconsisting of H, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, and C₃-C₆cycloalkyl; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, and C₃-C₆ cycloalkyl is independently optionallysubstituted by deuterium, fluoro, chloro, bromo, —OH, —CN, —OC₁-C₆alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, C₃-C₇ cycloalkyl, 3- to7-membered heterocycloalkyl, C₆-C₁₀ aryl, 5- to 7-membered heteroaryl,—CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), or—C(O)N(C₁-C₆ alkyl)₂;

R⁹ is H, fluoro, chloro, bromo, —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆ alkyl,—C(O)NH₂, —C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂;

R¹⁰ is H, fluoro, chloro or bromo; and

n is 1 or 2;

with the proviso that when R⁵ is H, R⁹ is selected from the groupconsisting of —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂.

In another aspect, the disclosure relates to a pharmaceuticalcomposition comprising a compound of the Formula I, or apharmaceutically acceptable salt thereof, and optionally at least one ormore of a pharmaceutically acceptable diluent, carrier or excipient.

In another aspect, the disclosure is directed to a method of treatingcancer in a patient comprising,

a. administering a therapeutically effective amount of a compound thatinhibits SRC and MET, and/or CSF1R. In some embodiments of this aspect,the compound that inhibits SRC and MET, and/or CSF1R is of the FormulaI. In some embodiments of this aspect, the cancer is gastric cancer,colon cancer, renal cancer, liver cancer, lung cancer, glioblastoma, orhead & neck cancer.

In another aspect, the disclosure is directed to a method of treatingcancer in a patient comprising,

a. administering a therapeutically effective amount of a compound thatinhibits SRC and MET, and/or CSF1R; and

b. administering a therapeutically effective amount of at least oneadditional anti-cancer agent. In some embodiments of this aspect, the atleast one additional anti-cancer agent is an EGFR inhibitor, or apharmaceutically acceptable salt thereof. In some embodiments of thisaspect, the additional anti-cancer agent is an antibody of EGFR. In someembodiments of this aspect, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments of this aspect,the cancer is gastric cancer, colon cancer, renal cancer, liver cancer,lung cancer, glioblastoma, or head & neck cancer.

In another aspect, the disclosure is directed to a compound thatinhibits SRC and MET, and/or CSF1R, or a pharmaceutically acceptablesalt thereof, for use in the treatment of cancer in a patient. In someembodiments of this aspect, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments of this aspect,the cancer is gastric cancer, colon cancer, renal cancer, liver cancer,lung cancer, glioblastoma, or head & neck cancer.

In another aspect, the disclosure is directed to a compound thatinhibits SRC and MET, and/or CSF1R, or a pharmaceutically acceptablesalt thereof, in combination with a therapeutically effective amount ofat least one additional anti-cancer agent, or a pharmaceuticallyacceptable salt thereof, for use in the treatment of cancer in apatient. In some embodiments of this aspect, the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof. In some embodiments of this aspect, the additionalanti-cancer agent is an antibody of EGFR. In some embodiments of thisaspect, the compound that inhibits SRC and MET, and/or CSF1R is of theFormula I. In some embodiments of this aspect, the cancer is gastriccancer, colon cancer, renal cancer, liver cancer, lung cancer,glioblastoma, or head & neck cancer.

In another aspect, the disclosure is directed to use of a compound thatinhibits SRC and MET, and/or CSF1R, or a pharmaceutically acceptablesalt thereof, for use in the treatment of cancer in a patient. In someembodiments of this aspect, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments of this aspect,the cancer is gastric cancer, colon cancer, renal cancer, liver cancer,lung cancer, glioblastoma, or head & neck cancer. In some embodiments ofthis aspect, the compound is administered in combination with atherapeutically effective amount of at least one additional anti-canceragent. In some embodiments of this aspect, the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof. In some embodiments of this aspect, the additionalanti-cancer agent is an antibody of EGFR.

In another aspect, the disclosure is directed to a compositioncomprising a compound that inhibits SRC and MET, and/or CSF1R, or apharmaceutically acceptable salt thereof, in a therapeutically effectiveamount, for use in the treatment of cancer in a patient. In someembodiments of this aspect, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments of this aspect,the cancer is gastric cancer, colon cancer, renal cancer, liver cancer,lung cancer, glioblastoma, or head & neck cancer. In some embodiments ofthis aspect, the compound is administered in combination with atherapeutically effective amount of at least one additional anti-canceragent. In some embodiments of this aspect, the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof. In some embodiments of this aspect, the additionalanti-cancer agent is an antibody of EGFR.

In yet another aspect, the disclosure relates to a synergisticcomposition of a compound that inhibits SRC and MET, and/or CSF1R, andan EGFR inhibitor, where the two components come into contact with eachother at a locus. In some embodiments of this aspect, the compound thatinhibits SRC and MET, and/or CSF1R is of the Formula I.

Additional embodiments, features, and advantages of the disclosure willbe apparent from the following detailed description and through practiceof the disclosure. The compounds of the present disclosure can bedescribed as embodiments in any of the following enumerated clauses. Itwill be understood that any of the embodiments described herein can beused in connection with any other embodiments described herein to theextent that the embodiments do not contradict one another.

1. A compound to of the Formula I

or a pharmaceutically acceptable salt thereof, wherein

X¹ and X² are independently —CR⁶R⁷—, S, S(O), S(O)₂, O or N(R⁸);

R¹ is H, deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, C₃-C₁₀ aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein each hydrogenatom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl andC₆-C₁₀ aryl is independently optionally substituted by deuterium,halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂,—NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆alkyl, —NHC(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂,—NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆alkyl), —NHS(O)₂(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl), —NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆alkyl)S(O)NH₂, —N(C₁-C₆ alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl),—NHS(O)₂NH(C₁-C₆ alkyl), —NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆alkyl)₂, —N(C₁-C₆ alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂, —SC₁-C₆ alkyl, —S(O)C₁-C₆alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆ alkyl), —S(O)₂NH(C₁-C₆ alkyl),—S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆ alkyl)₂, —P(C₁-C₆ alkyl)₂,—P(O)(C₁-C₆alkyl)₂, C₃-C₆ cycloalkyl, or 3- to 7-memberedheterocycloalkyl;

each R² and R³ is independently H, deuterium, C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁸ or—C(O)NR⁸R⁹; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl and C₆-C₁₀ aryl is independentlyoptionally substituted by deuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl,—NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂,—N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆alkyl)₂, —P(O)(C₁-C₆alkyl)₂, C₃-C₆ cycloalkyl, or 3- to7-membered heterocycloalkyl; or R² and R³ taken together with the carbonatoms to which they are attached optionally form a C₅-C₇ cycloalkyl or a5- to 7-membered heterocycloalkyl; or R² and R⁴ taken together with theatoms to which they are attached optionally form a 5- to 7-memberedheterocycloalkyl;

R⁴ is H, C₁-C₆ alkyl or 3- to 7-membered heterocycloalkyl, wherein eachhydrogen atom in C₁-C₆ alkyl or 3- to 7-membered heterocycloalkyl isindependently optionally substituted by halogen, —OH, —CN, —OC₁-C₆alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —CO₂H, —C(O)OC₁-C₆alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂, C₃-C₆cycloalkyl, or monocyclic 5- to 7-membered heterocycloalkyl;

R⁵ is H or —NR⁶R⁷;

each R⁶, R⁷ and R⁸ are each independently selected from the groupconsisting of H, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, and C₃-C₆cycloalkyl; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, and C₃-C₆ cycloalkyl is independently optionallysubstituted by deuterium, fluoro, chloro, bromo, —OH, —CN, —OC₁-C₆alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, C₃-C₇ cycloalkyl, 3- to7-membered heterocycloalkyl, C₆-C₁₀ aryl, 5- to 7-membered heteroaryl,—CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), or—C(O)N(C₁-C₆ alkyl)₂;

R⁹ is H, fluoro, chloro, bromo, —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆ alkyl,—C(O)NH₂, —C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂;

R¹⁰ is H, fluoro, chloro or bromo; and

n is 1 or 2;

with the proviso that when R⁵ is H, R⁹ is selected from the groupconsisting of —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂.

2. The compound of clause 1, or a pharmaceutically acceptable saltthereof, wherein R⁵ is H.

3. The compound of clause 2, or a pharmaceutically acceptable saltthereof, wherein R⁹ is —CN.

4. The compound of any of the preceding clauses, or a pharmaceuticallyacceptable salt thereof, wherein R¹⁰ is F.

5. The compound of clause 1, or a pharmaceutically acceptable saltthereof, wherein R⁵ is —NR⁶R⁷.

6. The compound of clause 5, or a pharmaceutically acceptable saltthereof, wherein R⁶ and R⁷ are H.

7. The compound of clause 5, or a pharmaceutically acceptable saltthereof, wherein R⁹ is —CN.

8. The compound of clause 6, or a pharmaceutically acceptable saltthereof, wherein R⁹ is —CN.

9. The compound of any one of clauses 5 to 8, or a pharmaceuticallyacceptable salt thereof, wherein R¹⁰ is fluoro.

10. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein X¹ is N(R⁸).

11. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R⁸ is C₁-C₆ alkyl,wherein each hydrogen atom is independently optionally substituted byfluoro, chloro, bromo, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl),

—N(C₁-C₆ alkyl)₂, C₃-C₇ cycloalkyl, 3- to 7-membered heterocycloalkyl,C₆-C₁₀ aryl, 5- to 7-membered heteroaryl, —CO₂H, —C(O)OC₁-C₆ alkyl,—C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), or —C(O)N(C₁-C₆ alkyl)₂.

12. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R⁸ is ethyl, propyl,iso-propyl, or methylcyclopropyl.

13. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein X² is O.

14. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R² is C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁷ or—C(O)NR⁷R⁸; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl and C₆-C₁₀ aryl is independentlyoptionally substituted by deuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl,—NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂,—N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆ alkyl)₂, —P(O)(C₁-C₆alkyl)², C₃-C₆ cycloalkyl, or 3-to 7-membered heterocycloalkyl, and R³ is H.

15. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R² is C₁-C₆ alkyl.

16. The compound of any one of the preceding clauses, or apharmaceutically acceptable salt thereof, wherein R² is methyl.

17. The compound of any one of clauses 1 to 14, or a pharmaceuticallyacceptable salt thereof, wherein R² is H, and R³ is H, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁷ or—C(O)NR⁷R⁸; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl, C₃-C₆ cycloalkyl and C₆-C₁₀ aryl is independentlyoptionally substituted by deuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl,—NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, N(C₁-C₆alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂,—N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆ alkyl)₂, —P(O)(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or 3-to 7-membered heterocycloalkyl.

18. The compound of any one of clauses 1 to 14, or a pharmaceuticallyacceptable salt thereof, wherein R² and R³ are H.

19. The compound of clause 1, selected from the group consisting of

or a pharmaceutically acceptable salt thereof.

20. A pharmaceutical composition comprising a compound of any one ofclauses 1 to 19, or a pharmaceutically acceptable salt thereof, and atleast one or more of a pharmaceutically acceptable diluent, carrier orexcipient.

21. A method of treating cancer in a patient comprising,

a. administering a therapeutically effective amount of a compound thatinhibits SRC and MET, and/or CSF1R.

22. The method of clause 22, wherein the compound that inhibits SRC andMET, and/or CSF1R is of the formula of any one of clauses 1 to 19.

23. The method of clause 21 or 22, wherein the cancer is gastric cancer,colon cancer, renal cancer, liver cancer, lung cancer, glioblastoma, orhead & neck cancer.

24. The method of any one of clauses 21 to 23, further comprising

b. administering a therapeutically effective amount of at least oneadditional anti-cancer agent.

25. The method of clause 24, wherein the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof.

26. The method of clause 24, wherein the additional anti-cancer agent isan antibody of EGFR.

27. The method of clause 26, wherein the antibody of EGFR is cetuximab,necitumumab or panitumumab.

28. The method of clause 24, wherein the additional anti-cancer agent isa small molecule inhibitor of EGFR.

29. The method of clause 28, wherein the small molecule inhibitor ofEGFR is afatinib, brigatinib, canertinib, dacomitinib, erlotinib,gefitinib, HKI 357, lapatinib, osimertinib, naquotinib, nazartinib,neratinib, olmutinib, pelitinib, PF-06747775, rociletinib, vandetanib,or a pharmaceutically acceptable salt thereof.

30. The method of any one of clauses 24, 28 or 29, wherein theadditional anti-cancer agent is gefitinib, or a pharmaceuticallyacceptable salt thereof.

31. The method of any one of clauses 24, 28 or 29, wherein theadditional anti-cancer agent is osimertinib, or a pharmaceuticallyacceptable salt thereof.

32. The method of any one of clauses 24, 28 or 29, wherein theadditional anti-cancer agent is erlotinib, or a pharmaceuticallyacceptable salt thereof.

33. A compound that inhibits SRC and MET, and/or CSF1R, or apharmaceutically acceptable salt thereof, for use in the treatment ofcancer in a patient.

34. The compound of clause 33, wherein the compound that inhibits SRCand MET, and/or CSF1R is of the formula of any one of clauses 1 to 19.

35. The compound of clause 33 or 34, wherein the cancer is gastriccancer, colon cancer, renal cancer, liver cancer, lung cancer,glioblastoma, or head & neck cancer.

36. The compound of any one of clauses 33 to 35, in combination with atherapeutically effective amount of at least one additional anti-canceragent.

37. The compound of clause 36, wherein the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof.

38. The compound of clause 36, wherein the additional anti-cancer agentis an antibody of EGFR.

39. The compound of clause 38, wherein the antibody of EGFR iscetuximab, necitumumab or panitumumab.

40. The compound of clause 36, wherein the additional anti-cancer agentis a small molecule inhibitor of EGFR.

41. The compound of clause 40, wherein the small molecule inhibitor ofEGFR is afatinib, brigatinib, canertinib, dacomitinib, erlotinib,gefitinib, HKI 357, lapatinib, osimertinib, naquotinib, nazartinib,neratinib, olmutinib, pelitinib, PF-06747775, rociletinib, vandetanib,or a pharmaceutically acceptable salt thereof.

42. The compound of any one of clauses 36, 40 or 41, wherein theadditional anti-cancer agent is gefitinib, or a pharmaceuticallyacceptable salt thereof.

43. The compound of any one of clauses 36, 40 or 41, wherein theadditional anti-cancer agent is osimertinib, or a pharmaceuticallyacceptable salt thereof.

44. The compound of any one of clauses 36, 40 or 41, wherein theadditional anti-cancer agent is erlotinib, or a pharmaceuticallyacceptable salt thereof.

45. Use of a compound that inhibits SRC and MET, and/or CSF1R, or apharmaceutically acceptable salt thereof, in the preparation of amedicament for use in the treatment of cancer.

46. The use of clause 45, wherein the compound that inhibits SRC andMET, and/or CSF1R is of the formula of any one of clauses 1 to 19.

47. The use of clause 45 or 46, wherein the cancer is gastric cancer,colon cancer, renal cancer, liver cancer, lung cancer, glioblastoma, orhead & neck cancer.

48. The use of any one of clauses 45 to 47, in combination with atherapeutically effective amount of at least one additional anti-canceragent.

49. The use of clause 48, wherein the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof.

50. The use of clause 48, wherein the additional anti-cancer agent is anantibody of EGFR.

51. The use of clause 50, wherein the antibody of EGFR cetuximab,necitumumab or panitumumab.

52. The use of clause 48, wherein the additional anti-cancer agent is asmall molecule inhibitor of EGFR.

53. The use of clause 52, wherein the small molecule inhibitor of EGFRis afatinib, brigatinib, canertinib, dacomitinib, erlotinib, gefitinib,HKI 357, lapatinib, osimertinib, naquotinib, nazartinib, neratinib,olmutinib, pelitinib. PF-06747775, rociletinib, vandetanib, or apharmaceutically acceptable salt thereof.

54. The use of any one of clauses 48, 52 or 53, wherein the additionalanti-cancer agent is gefitinib, or a pharmaceutically acceptable saltthereof.

55. The use of any one of clauses 48, 52 or 53, wherein the additionalanti-cancer agent is osimertinib, or a pharmaceutically acceptable saltthereof.

56. The use of any one of clauses 48, 52 or 53, wherein the additionalanti-cancer agent is erlotinib, or a pharmaceutically acceptable saltthereof.

57. A composition comprising a compound that inhibits SRC and MET,and/or CSF1R, or a pharmaceutically acceptable salt thereof, in atherapeutically effective amount, for use in the treatment of cancer ina patient.

58. The composition of clause 57, wherein the compound that inhibits SRCand MET, and/or CSF1R is of the formula of any one of clauses 1 to 20.

59. The composition of clause 56 or 57, wherein the cancer is gastriccancer, colon cancer, renal cancer, liver cancer, lung cancer,glioblastoma, or head & neck cancer.

60. The composition of any one of clauses 57 to 59, in combination witha therapeutically effective amount of at least one additionalanti-cancer agent.

61. The composition of clause 60, wherein the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof.

62. The composition of clause 60, wherein the additional anti-canceragent is an antibody of EGFR.

63. The composition of clause 62, wherein the antibody of EGFRcetuximab, necitumumab or panitumumab.

64. The composition of clause 60, wherein the additional anti-canceragent is a small molecule inhibitor of EGFR.

65. The composition of clause 64, wherein the small molecule inhibitorof EGFR is afatinib, brigatinib, canertinib, dacomitinib, erlotinib,gefitinib, HKI 357, lapatinib, osimertinib, naquotinib, nazartinib,neratinib, olmutinib, pelitinib, PF-06747775, rociletinib, vandetanib,or a pharmaceutically acceptable salt thereof.

66. The composition of any one of clauses 60, 64 or 65, wherein theadditional anti-cancer agent is gefitinib, or a pharmaceuticallyacceptable salt thereof.

67. The composition of any one of clauses 60, 64 or 65, wherein theadditional anti-cancer agent is osimertinib, or a pharmaceuticallyacceptable salt thereof.

68. The composition of any one of clauses 60, 64 or 65, wherein theadditional anti-cancer agent is erlotinib, or a pharmaceuticallyacceptable salt thereof.

69. A synergistic composition of a compound that inhibits SRC and MET,and/or CSF1R, and an EGFR inhibitor, where the two components come intocontact with each other at a locus.

70. The synergistic composition of clause 69, wherein the compound thatinhibits SRC and MET, and/or CSF1R is of the formula of any one ofclauses 1 to 19.

71. The synergistic composition of clause 69 or 70, wherein the locus isa patient.

72. The synergistic composition of clause 69 or 70, wherein the locus isa cancer.

73. The synergistic composition of clause 72, wherein the cancer isgastric cancer, colon cancer, renal cancer, liver cancer, lung cancer,glioblastoma, or head & neck cancer.

74. The synergistic composition of any one of clauses 69 to 73, whereinthe EGFR inhibitor is an antibody of EGFR.

75. The synergistic composition of clause 74, wherein the antibody ofEGFR cetuximab, necitumumab or panitumumab.

76. The synergistic composition of any one of clauses 69 to 73, whereinthe EGFR inhibitor is a small molecule inhibitor of EGFR.

77. The synergistic composition of clause 76, wherein the small moleculeinhibitor of EGFR is afatinib, brigatinib, canertinib, dacomitinib,erlotinib, gefitinib, HKI 357, lapatinib, osimertinib, naquotinib,nazartinib, neratinib, olmutinib, pelitinib, PF-06747775, rociletinib,vandetanib, or pharmaceutically acceptable salts thereof.

78. The synergistic composition of any one of clauses 69-73, 76 or 77,wherein the EGFR inhibitor is gefitinib, or a pharmaceuticallyacceptable salt thereof.

79. The synergistic composition of any one of clauses 69-73, 76 or 77,wherein the EGFR inhibitor is osimertinib, or a pharmaceuticallyacceptable salt thereof.

80. The synergistic composition of any one of clauses 69-73, 76 or 77,wherein the EGFR inhibitor is erlotinib, or a pharmaceuticallyacceptable salt thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a gel image of studies of MET phosphorylation in SNU-5cells after 4 hour incubation with Compound 5. The gel shows thatCompound 5 inhibited MET phosphorylation in SNU-5 cells.

FIG. 2 shows a gel image of studies of phosphorylation of MET anddownstream effectors in MKN-45 cells after 16 hour incubation withCompound 5. The gel shows that Compound 5 inhibited MET phosphorylationand downstream effectors in MKN-45 cells.

FIG. 3 is a graph showing the effects of Compound 5, capmatinib, andAZD9291 on HCC827 cell proliferation. A strong synergistic activity wasobserved in the combination of AZD9291 with Compound 5 with an IC₅₀ of 2nM and Emax 71% in HCC827 cell proliferation assay. (▾) capmatinib(IC₅₀: >10000 nM, Emax %: -), (▴) Compound 5 (IC₅₀: 3000 nM, Emax %: -),(▪) AZD9291 (IC₅₀: 5 nM (partial), Emax %: 47), (♦) capmatinib(1+AZD9291 (IC₅₀: 5 nM (partial), Emax %: 47), (•) Compound 5 (1+AZD9291(IC₅₀: 2 nM, Emax %: 71).

FIG. 4 shows the effects of Compound 5, capmatinib, AZD9291 andcombinations on the apoptosis of HCC827 cells after 48 hour incubation.Compound 5 synergized with AZD9291 for apoptosis in HCC827 cell line.

FIG. 5 shows a wound healing assay in which Compound 5 and capmatinibinhibited cell migration of MKN-45 cells.

FIG. 6 shows a wound healing assay in which Compound 5 inhibited cellmigration of HCC827, and campmatinib showed a minimal effect.

FIG. 7 is a graph showing the effect of Compound 5 on tumor growth inthe MKN-45 xenograft model. (•) vehicle, (▪) Compound 5 at 3 mg/kg BID,(▴) Compound 5 at 10 mg/kg BID, (▾) Compound 5 at 30 mg/kg BID.

FIG. 8 shows the effect of Compound 5 on the body weight of mice bearingMKN-45 xenograft tumors. (•) vehicle, (▪) Compound 5 at 3 mg/kg BID, (▴)Compound 5 at 10 mg/kg BID, (▾) Compound 5 at 30 mg/kg BID.

FIG. 9 shows a gel image of studies of inhibition of MET phosphorylationby Compound 5 in the MKN-45 xenograft model.

FIG. 10 is a chart showing the effect of Compound 5 on thephosphorylation od Met Y1234/1235 in MKN-45 tumors. (•) Vehicle, (▪)Compound 5 at 10 mg/kg 4 Hrs, (▴) Compound 5 at 10 mg/kg 12 Hrs, (▾)Compound 5 at 3 mg/kg 4 Hrs, (♦) Compound 5 at 3 mg/kg 12 Hrs.

FIG. 11 is a chart showing the anti-tumor activity of Compound 5 inLU2503 PDX tumors. (•) Vehicle, (▪) Compound 5 at 15 mg/kg BID.

FIG. 12 is a chart showing the body weights of mice bearing LU2503 PDXtumors treated with Compound 5. (•) Vehicle, (▪) Compound 5 at 15 mg/kgBID.

FIG. 13 is a chart showing the anti-tumor activity of Compound 5 in BaF3ETV6-CSF1R tumors. (•) Vehicle, (▪) Compound 5 at 5 mg/kg BID, (▴)Compound 5 at 15 mg/kg BID.

FIG. 14 is a chart showing the body weights of mice bearing LU25BaF3ETV6-CSF1R tumors treated with Compound 5. (•) Vehicle, (▪) Compound 5at 5 mg/kg BID, (▴) Compound 5 at 15 mg/kg BID.

FIG. 15 is a chart showing the anti-tumor activity of Compound 5 in MC38synergistic mouse tumor model. (•) Vehicle, (▪) Compound 5 at 15 mg/kgBID.

FIG. 16 is a chart showing the body weights of mice bearing MC38synergistic mouse tumor model treated with Compound 5. (•) Vehicle, (▪)Compound 5 at 15 mg/kg BID.

FIG. 17A-17G are graphs showing FACS analysis of tumor samples from eachgroup after Day 7 treatment with Compound 5. FIG. 17A shows % in CD45+cells; CD8 T-cells. FIG. 17B shows % in CD45+ cells; CD4 T-cells. FIG.17C shows % in CD45+ cells; T-Reg. FIG. 17D shows % in CD45+ cells;MDSCs. FIG. 17E shows % in CD45+ cells; TAMs. FIG. 17F shows % in CD45+cells; M1 macrophage. 17G shows % in CD45+ cells; M2 macrophage.

FIG. 18A-18G are graphs showing FACS analysis of tumor samples from eachgroup after Day 11 treatment with Compound 5. FIG. 18A shows % in CD45+cells; CD4 T-cells. FIG. 18B shows % in CD45+ cells; CD8 T-cells. FIG.18C shows % in CD45+ cells; T-Reg. FIG. 18D shows % in CD45+ cells;MDSCs. FIG. 18E shows % in CD45+ cells; TAMs. FIG. 18F shows % in CD45+cells; M1 macrophage. 18G shows % in CD45+ cells; M2 macrophage.

FIG. 19 is a chart showing the in-vivo efficacy of Compound 5 insubcutaneous MC38 synergistic mouse tumor model. (•) G1-Vehiclie+ISOIiG; (▪) Compound 5; (▴) Anti-PD-1, (▾) Compound 5+Anti-PD-1.

FIG. 20 is a chart showing the body weights of mice bearing subcutaneousMC38 synergistic mouse tumor model. (•) G1-Vehiclie+ISO IiG; (▪)Compound 5; (▴) Anti-PD-1, (▾) Compound 5+Anti-PD-1.

DETAILED DESCRIPTION

Before the present disclosure is further described, it is to beunderstood that this disclosure is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present disclosure will be limited only by the appendedclaims.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this disclosure belongs. All patents, applications,published applications and other publications referred to herein areincorporated by reference in their entireties. If a definition set forthin this section is contrary to or otherwise inconsistent with adefinition set forth in a patent, application, or other publication thatis herein incorporated by reference, the definition set forth in thissection prevails over the definition incorporated herein by reference.

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include plural referents unless the context clearly dictatesotherwise. It is further noted that the claims may be drafted to excludeany optional element. As such, this statement is intended to serve asantecedent basis for use of such exclusive terminology as “solely,”“only” and the like in connection with the recitation of claim elements,or use of a “negative” limitation.

As used herein, the terms “including,” “containing,” and “comprising”are used in their open, non-limiting sense.

To provide a more concise description, some of the quantitativeexpressions given herein are not qualified with the term “about”. It isunderstood that, whether the term “about” is used explicitly or not,every quantity given herein is meant to refer to the actual given value,and it is also meant to refer to the approximation to such given valuethat would reasonably be inferred based on the ordinary skill in theart, including equivalents and approximations due to the experimentaland/or measurement conditions for such given value. Whenever a yield isgiven as a percentage, such yield refers to a mass of the entity forwhich the yield is given with respect to the maximum amount of the sameentity that could be obtained under the particular stoichiometricconditions. Concentrations that are given as percentages refer to massratios, unless indicated differently.

Except as otherwise noted, the methods and techniques of the presentembodiments are generally performed according to conventional methodswell known in the art and as described in various general and morespecific references that are cited and discussed throughout the presentspecification. See, e.g., Loudon, Organic Chemistry, Fourth Edition, NewYork: Oxford University Press, 2002, pp. 360-361, 1084-1085; Smith andMarch, March's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, Fifth Edition, Wiley-Interscience, 2001.

Chemical nomenclature for compounds described herein has generally beenderived using the commercially-available ACD/Name 2014 (ACD/Labs) orChemBioDraw Ultra 13.0 (Perkin Elmer).

It is appreciated that certain features of the disclosure, which are,for clarity, described in the context of separate embodiments, may alsobe provided in combination in a single embodiment. Conversely, variousfeatures of the disclosure, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination. All combinations of the embodimentspertaining to the chemical groups represented by the variables arespecifically embraced by the present disclosure and are disclosed hereinjust as if each and every combination was individually and explicitlydisclosed, to the extent that such combinations embrace compounds thatare stable compounds (i.e., compounds that can be isolated,characterized, and tested for biological activity). In addition, allsubcombinations of the chemical groups listed in the embodimentsdescribing such variables are also specifically embraced by the presentdisclosure and are disclosed herein just as if each and every suchsub-combination of chemical groups was individually and explicitlydisclosed herein.

Definitions

As used herein, the term “alkyl” includes a chain of carbon atoms, whichis optionally branched and contains from 1 to 20 carbon atoms. It is tobe further understood that in certain embodiments, alkyl may beadvantageously of limited length, including C₁-C₁₂, C₁-C₁₀, C₁-C₉,C₁-C₈, C₁-C₇, C₁-C₆, and C₁-C₄, Illustratively, such particularlylimited length alkyl groups, including C₁-C₈, C₁-C₇, C₁-C₆, and C₁-C₄,and the like may be referred to as “lower alkyl.” Illustrative alkylgroups include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, 2-pentyl,3-pentyl, neopentyl, hexyl, heptyl, octyl, and the like. Alkyl may besubstituted or unsubstituted. Typical substituent groups includecycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy,mercapto, alkylthio, arylthio, cyano, halo, carbonyl, oxo, (═O),thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl,C-amido, N-amido, C-carboxy, O-carboxy, nitro, and amino, or asdescribed in the various embodiments provided herein. It will beunderstood that “alkyl” may be combined with other groups, such as thoseprovided above, to form a functionalized alkyl. By way of example, thecombination of an “alkyl” group, as described herein, with a “carboxy”group may be referred to as a “carboxyalkyl” group. Other non-limitingexamples include hydroxyalkyl, aminoalkyl, and the like.

As used herein, the term “alkenyl” includes a chain of carbon atoms,which is optionally branched, and contains from 2 to 20 carbon atoms,and also includes at least one carbon-carbon double bond (i.e. C═C). Itwill be understood that in certain embodiments, alkenyl may beadvantageously of limited length, including C₂-C₁₂, C₂-C₉, C₂-C₈, C₂-C₇,C₂-C₆, and C₂-C₄. Illustratively, such particularly limited lengthalkenyl groups, including C₂-C₈, C₂-C₇, C₂-C₆, and C₂-C₄ may be referredto as lower alkenyl. Alkenyl may be unsubstituted, or substituted asdescribed for alkyl or as described in the various embodiments providedherein. Illustrative alkenyl groups include, but are not limited to,ethenyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, and the like.

As used herein, the term “alkynyl” includes a chain of carbon atoms,which is optionally branched, and contains from 2 to 20 carbon atoms,and also includes at least one carbon-carbon triple bond (i.e. C≡C). Itwill be understood that in certain embodiments, alkynyl may each beadvantageously of limited length, including C₂-C₁₂, C₂-C₉, C₂-C₈, C₂-C₇,C₂-C₆, and C₂-C₄. Illustratively, such particularly limited lengthalkynyl groups, including C₂-C₈, C₂-C₇, C₂-C₆, and C₂-C₄ may be referredto as lower alkynyl. Alkenyl may be unsubstituted, or substituted asdescribed for alkyl or as described in the various embodiments providedherein. Illustrative alkenyl groups include, but are not limited to,ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, and the like.

As used herein, the term “aryl” refers to an all-carbon monocyclic orfused-ring polycyclic groups of 6 to 12 carbon atoms having a completelyconjugated pi-electron system. It will be understood that in certainembodiments, aryl may be advantageously of limited size such as C₆-C₁₀aryl. Illustrative aryl groups include, but are not limited to, phenyl,naphthylenyl and anthracenyl. The aryl group may be unsubstituted, orsubstituted as described for alkyl or as described in the variousembodiments provided herein.

As used herein, the term “cycloalkyl” refers to a 3 to 15 memberall-carbon monocyclic ring, including an all-carbon 5-member/6-member or6-member/6-member fused bicyclic ring, or a multicyclic fused ring (a“fused” ring system means that each ring in the system shares anadjacent pair of carbon atoms with each other ring in the system) group,where one or more of the rings may contain one or more double bonds butthe cycloalkyl does not contain a completely conjugated pi-electronsystem. It will be understood that in certain embodiments, cycloalkylmay be advantageously of limited size such as C₃-C₁₃, C₃-C₉, C₃-C₆ andC₄-C₆. Cycloalkyl may be unsubstituted, or substituted as described foralkyl or as described in the various embodiments provided herein.Illustrative cycloalkyl groups include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclopentadienyl,cyclohexyl, cyclohexenyl, cycloheptyl, adamantyl, norbornyl,norbornenyl, 9H-fluoren-9-yl, and the like. Illustrative examples ofcycloalkyl groups shown in graphical representations include thefollowing entities, in the form of properly bonded moieties:

As used herein, the term “heterocycloalkyl” refers to a monocyclic orfused ring group having in the ring(s) from 3 to 12 ring atoms, in whichat least one ring atom is a heteroatom, such as nitrogen, oxygen orsulfur, the remaining ring atoms being carbon atoms. Heterocycloalkylmay optionally contain 1, 2, 3 or 4 heteroatoms. Heterocycloalkyl mayalso have one of more double bonds, including double bonds to nitrogen(e.g. C═N or N═N) but does not contain a completely conjugatedpi-electron system. It will be understood that in certain embodiments,heterocycloalkyl may be advantageously of limited size such as 3- to7-membered heterocycloalkyl, 5- to 7-membered heterocycloalkyl, and thelike. Heterocycloalkyl may be unsubstituted, or substituted as describedfor alkyl or as described in the various embodiments provided herein.Illustrative heterocycloalkyl groups include, but are not limited to,oxiranyl, thianaryl, azetidinyl, oxetanyl, tetrahydrofuranyl,pyrrolidinyl, tetrahydropyranyl, piperidinyl, 1,4-dioxanyl, morpholinyl,1,4-dithianyl, piperazinyl, oxepanyl, 3,4-dihydro-2H-pyranyl,5,6-dihydro-2H-pyranyl, 2H-pyranyl, 1, 2, 3, 4-tetrahydropyridinyl, andthe like. Illustrative examples of heterocycloalkyl groups shown ingraphical representations include the following entities, in the form ofproperly bonded moieties:

As used herein, the term “heteroaryl” refers to a monocyclic or fusedring group of 5 to 12 ring atoms containing one, two, three or four ringheteroatoms selected from nitrogen, oxygen and sulfur, the remainingring atoms being carbon atoms, and also having a completely conjugatedpi-electron system. It will be understood that in certain embodiments,heteroaryl may be advantageously of limited size such as 3- to7-membered heteroaryl, 5- to 7-membered heteroaryl, and the like.Heteroaryl may be unsubstituted, or substituted as described for alkylor as described in the various embodiments provided herein. Illustrativeheteroaryl groups include, but are not limited to, pyrrolyl, furanyl,thiophenyl, imidazolyl, oxazolyl, thiazolyl, pyrazolyl, pyridinyl,pyrimidinyl, quinolinyl, isoquinolinyl, purinyl, tetrazolyl, triazinyl,pyrazinyl, tetrazinyl, quinazolinyl, quinoxalinyl, thienyl, isoxazolyl,isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzimidazolyl,benzoxazolyl, benzthiazolyl, benzisoxazolyl, benzisothiazolyl andcarbazoloyl, and the like. Illustrative examples of heteroaryl groupsshown in graphical representations, include the following entities, inthe form of properly bonded moieties:

As used herein, “hydroxy” or “hydroxyl” refers to an —OH group.

As used herein, “alkoxy” refers to both an —O-(alkyl) or an—O-(unsubstituted cycloalkyl) group. Representative examples include,but are not limited to, methoxy, ethoxy, propoxy, butoxy,cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and thelike.

As used herein, “aryloxy” refers to an —O-aryl or an —O-heteroarylgroup. Representative examples include, but are not limited to, phenoxy,pyridinyloxy, furanyloxy, thienyloxy, pyrimidinyloxy, pyrazinyloxy, andthe like.

As used herein, “mercapto” refers to an —SH group.

As used herein, “alkylthio” refers to an —S-(alkyl) or an—S-(unsubstituted cycloalkyl) group. Representative examples include,but are not limited to, methylthio, ethylthio, propylthio, butylthio,cyclopropylthio, cyclobutylthio, cyclopentylthio, cyclohexylthio, andthe like.

As used herein, “arylthio” refers to an —S-aryl or an —S-heteroarylgroup. Representative examples include, but are not limited to,phenylthio, pyridinylthio, furanylthio, thienylthio, pyrimidinylthio,and the like.

As used herein, “halo” or “halogen” refers to fluorine, chlorine,bromine or iodine.

As used herein, “cyano” refers to a —CN group.

The term “oxo” represents a carbonyl oxygen. For example, a cyclopentylsubstituted with oxo is cyclopentanone.

As used herein, “bond” refers to a covalent bond.

The term “substituted” means that the specified group or moiety bearsone or more substituents. The term “unsubstituted” means that thespecified group bears no substituents. Where the term “substituted” isused to describe a structural system, the substitution is meant to occurat any valency-allowed position on the system. In some embodiments,“substituted” means that the specified group or moiety bears one, two,or three substituents. In other embodiments, “substituted” means thatthe specified group or moiety bears one or two substituents. In stillother embodiments, “substituted” means the specified group or moietybears one substituent.

As used herein, “optional” or “optionally” means that the subsequentlydescribed event or circumstance may but need not occur, and that thedescription includes instances where the event or circumstance occursand instances in which it does not. For example, “wherein each hydrogenatom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3-to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclicheteroaryl is independently optionally substituted by C₁-C₆ alkyl” meansthat an alkyl may be but need not be present on any of the C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, or mono- or bicyclic heteroaryl byreplacement of a hydrogen atom for each alkyl group, and the descriptionincludes situations where the C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, ormono- or bicyclic heteroaryl is substituted with an alkyl group andsituations where the C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, or mono- orbicyclic heteroaryl is not substituted with the alkyl group.

As used herein, “independently” means that the subsequently describedevent or circumstance is to be read on its own relative to other similarevents or circumstances. For example, in a circumstance where severalequivalent hydrogen groups are optionally substituted by another groupdescribed in the circumstance, the use of “independently optionally”means that each instance of a hydrogen atom on the group may besubstituted by another group, where the groups replacing each of thehydrogen atoms may be the same or different. Or for example, wheremultiple groups exist all of which can be selected from a set ofpossibilities, the use of “independently” means that each of the groupscan be selected from the set of possibilities separate from any othergroup, and the groups selected in the circumstance may be the same ordifferent.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which counter ions which may be used in pharmaceuticals.See, generally, S. M. Berge, et al., “Pharmaceutical Salts,” J. Pharm.Sci., 1977, 66, 1-19. Preferred pharmaceutically acceptable salts arethose that are pharmacologically effective and suitable for contact withthe tissues of subjects without undue toxicity, irritation, or allergicresponse. A compound described herein may possess a sufficiently acidicgroup, a sufficiently basic group, both types of functional groups, ormore than one of each type, and accordingly react with a number ofinorganic or organic bases, and inorganic and organic acids, to form apharmaceutically acceptable salt. Such salts include:

(1) acid addition salts, which can be obtained by reaction of the freebase of the parent compound with inorganic acids such as hydrochloricacid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, andperchloric acid and the like, or with organic acids such as acetic acid,oxalic acid, (D) or (L) malic acid, maleic acid, methane sulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaricacid, citric acid, succinic acid or malonic acid and the like; or

(2) salts formed when an acidic proton present in the parent compoundeither is replaced by a metal ion, e.g., an alkali metal ion, analkaline earth ion, or an aluminum ion; or coordinates with an organicbase such as ethanolamine, diethanolamine, triethanolamine,trimethamine, N-methylglucamine, and the like.

Pharmaceutically acceptable salts are well known to those skilled in theart, and any such pharmaceutically acceptable salt may be contemplatedin connection with the embodiments described herein. Examples ofpharmaceutically acceptable salts include sulfates, pyrosulfates,bisulfates, sulfites, bisulfites, phosphates, monohydrogen-phosphates,dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides,bromides, iodides, acetates, propionates, decanoates, caprylates,acrylates, formates, isobutyrates, caproates, heptanoates, propiolates,oxalates, malonates, succinates, suberates, sebacates, fumarates,maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates,chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates,methoxybenzoates, phthalates, sulfonates, methylsulfonates,propylsulfonates, besylates, xylenesulfonates, naphthalene-1-sulfonates,naphthalene-2-sulfonates, phenylacetates, phenylpropionates,phenylbutyrates, citrates, lactates, γ-hydroxybutyrates, glycolates,tartrates, and mandelates. Lists of other suitable pharmaceuticallyacceptable salts are found in Remington's Pharmaceutical Sciences, 17thEdition, Mack Publishing Company, Easton, Pa., 1985.

For a compound of Formula I that contains a basic nitrogen, apharmaceutically acceptable salt may be prepared by any suitable methodavailable in the art, for example, treatment of the free base with aninorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuricacid, sulfamic acid, nitric acid, boric acid, phosphoric acid, and thelike, or with an organic acid, such as acetic acid, phenylacetic acid,propionic acid, stearic acid, lactic acid, ascorbic acid, maleic acid,hydroxymaleic acid, isethionic acid, succinic acid, valeric acid,fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid,salicylic acid, oleic acid, palmitic acid, lauric acid, a pyranosidylacid, such as glucuronic acid or galacturonic acid, an alpha-hydroxyacid, such as mandelic acid, citric acid, or tartaric acid, an aminoacid, such as aspartic acid or glutamic acid, an aromatic acid, such asbenzoic acid, 2-acetoxybenzoic acid, naphthoic acid, or cinnamic acid, asulfonic acid, such as laurylsulfonic acid, p-toluenesulfonic acid,methanesulfonic acid, or ethanesulfonic acid, or any compatible mixtureof acids such as those given as examples herein, and any other acid andmixture thereof that are regarded as equivalents or acceptablesubstitutes in light of the ordinary level of skill in this technology.

The disclosure also relates to pharmaceutically acceptable prodrugs ofthe compounds of Formula I, and treatment methods employing suchpharmaceutically acceptable prodrugs. The term “prodrug” means aprecursor of a designated compound that, following administration to asubject, yields the compound in vivo via a chemical or physiologicalprocess such as solvolysis or enzymatic cleavage, or under physiologicalconditions (e.g., a prodrug on being brought to physiological pH isconverted to the compound of Formula I). A “pharmaceutically acceptableprodrug” is a prodrug that is non-toxic, biologically tolerable, andotherwise biologically suitable for administration to the subject.Illustrative procedures for the selection and preparation of suitableprodrug derivatives are described, for example, in “Design of Prodrugs”,ed. H. Bundgaard, Elsevier, 1985.

The present disclosure also relates to pharmaceutically activemetabolites of compounds of Formula I, and uses of such metabolites inthe methods of the disclosure. A “pharmaceutically active metabolite”means a pharmacologically active product of metabolism in the body of acompound of Formula I, or pharmaceutically acceptable salt thereof.Prodrugs and active metabolites of a compound may be determined usingroutine techniques known or available in the art. See, e.g., Bertoliniet al., J. Med. Chem. 1997, 40, 2011-2016; Shan et al., J. Pharm. Sci.1997, 86 (7), 765-767; Bagshawe, Drug Dev. Res. 1995, 34, 220-230;Bodor, Adv. Drug Res. 1984, 13, 255-331; Bundgaard, Design of Prodrugs(Elsevier Press, 1985); and Larsen, Design and Application of Prodrugs,Drug Design and Development (Krogsgaard-Larsen et al., eds., HarwoodAcademic Publishers, 1991).

Any formula depicted herein is intended to represent a compound of thatstructural formula as well as certain variations or forms. For example,a formula given herein is intended to include a racemic form, or one ormore enantiomeric, diastereomeric, or geometric isomers, or a mixturethereof. Additionally, any formula given herein is intended to referalso to a hydrate, solvate, or polymorph of such a compound, or amixture thereof. For example, it will be appreciated that compoundsdepicted by a structural formula containing the symbol “

” include both stereoisomers for the carbon atom to which the symbol “

” is attached, specifically both the bonds “

” and “

” are encompassed by the meaning of “

”. For example, in some exemplary embodiments, certain compoundsprovided herein can be described by the formula

which formula will be understood to encompass compounds having bothstereochemical configurations at the relevant carbon atom. Specifically,in this example, the configurations can be described by the formulas

Any formula given herein is also intended to represent unlabeled formsas well as isotopically labeled forms of the compounds. Isotopicallylabeled compounds have structures depicted by the formulas given hereinexcept that one or more atoms are replaced by an atom having a selectedatomic mass or mass number. Examples of isotopes that can beincorporated into compounds of the disclosure include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, andiodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S,¹⁸F, ³⁶Cl, and ¹²⁵I, respectively. Such isotopically labelled compoundsare useful in metabolic studies (preferably with ¹⁴C), reaction kineticstudies (with, for example ²H or ³H), detection or imaging techniques[such as positron emission tomography (PET) or single-photon emissioncomputed tomography (SPECT)] including drug or substrate tissuedistribution assays, or in radioactive treatment of patients. Further,substitution with heavier isotopes such as deuterium (i.e., ²H) mayafford certain therapeutic advantages resulting from greater metabolicstability, for example increased in vivo half-life or reduced dosagerequirements. Isotopically labeled compounds of this disclosure andprodrugs thereof can generally be prepared by carrying out theprocedures disclosed in the schemes or in the examples and preparationsdescribed below by substituting a readily available isotopically labeledreagent for a non-isotopically labeled reagent.

Any disubstituent referred to herein is meant to encompass the variousattachment possibilities when more than one of such possibilities areallowed. For example, reference to disubstituent -A-B—, where A≠B,refers herein to such disubstituent with A attached to a firstsubstituted member and B attached to a second substituted member, and italso refers to such disubstituent with A attached to the secondsubstituted member and B attached to the first substituted member. Theuse of “- - -” in connection with the various chemical formulae providedherein to describe the various embodiments refers to a covalent bond(also referred to as a point of attachment) from the group to which “- --” to the remainder of the molecule.

Representative Embodiments

In some embodiments, compounds described herein comprise a moiety of theformula

wherein R⁵ is —NR⁶R⁷; and R⁶ and R⁷ are each independently selected fromthe group consisting of H, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,and C₃-C₆ cycloalkyl; wherein each hydrogen atom in C₁-C₆ alkyl, C₂-C₆alkenyl, C₂-C₆ alkynyl, and C₃-C₆ cycloalkyl is independently optionallysubstituted by fluoro, chloro, bromo, —OH, —CN, —OC₁-C₆ alkyl, —NH₂,—NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, C₃-C₇ cycloalkyl, 3- to 7-memberedheterocycloalkyl, C₆-C₁₀ aryl, 5- to 7-membered heteroaryl, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), or —C(O)N(C₁-C₆alkyl)₂.

In some embodiments, compounds described herein comprise a moiety of theformula

In still other embodiments, compounds described herein comprise a moietyof the formula

wherein R⁹ is H, fluoro, chloro, bromo, —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂; and R¹⁰is H, fluoro, chloro or bromo.

In still other embodiments, compounds described herein comprise a moietyof the formula

wherein R⁹ is fluoro, chloro, bromo, —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂; and R¹⁰is fluoro, chloro or bromo.

In some embodiments, when compounds described herein comprise a moietyof the formula

then R⁹ in the moiety of the formula

then R⁹ is selected from the group consisting of —CN, —CF₃, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆alkyl)₂.

In still other embodiments, compounds described herein comprise a moietyof the formula

In still other embodiments, compounds described herein comprise a moietyof the formula

and a moiety of the formula

In still other embodiments, compounds described herein comprise a moietyof the formula

and a moiety of the formula

In still other embodiments, compounds described herein comprise a moietyof the formula

and a moiety of the formula

In still other embodiments, compounds described herein comprise a moietyof the formula

and a moiety of the formula

In some embodiments, X¹ is —N(R⁸)—. In some embodiments, X² is —O—. Insome embodiments, X¹ is —N(R⁸)—, and X² is —O—.

In some embodiments, R¹ is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, C₃-C₁₀ aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein eachhydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl and C₆-C₁₀ aryl is independently optionally substituted bydeuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl,—NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆ alkyl)₂, —P(O)(C₁-C₆alkyl)₂, C₃-C₆ cycloalkyl, or 3-to 7-membered heterocycloalkyl. In some embodiments, R¹ is H.

In some embodiments, R² is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein eachhydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl and C₆-C₁₀ aryl is independently optionally substituted bydeuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl,—NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆alkyl)₂, —P(O)(C₁-C₆alkyl)₂, C₃-C₆ cycloalkyl, or 3- to7-membered heterocycloalkyl.

In some embodiments, R² is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,C₃-C₆ cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein eachhydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl and C₆-C₁₀ aryl is independently optionally substituted bydeuterium, halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl,—NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆ alkyl),—NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆ alkyl)₂, —P(O)(C₁-C₆alkyl)₂, C₃-C₆ cycloalkyl, or 3-to 7-membered heterocycloalkyl, and R³ is H.

In some embodiments, R² is C₁-C₆ alkyl, wherein each hydrogen atom inC₁-C₆ alkyl is independently optionally substituted with one or moremoieties selected from group consisting of —F, —OH, —OC₁-C₆ alkyl, —NH₂,—NH(C₁-C₆ alkyl), and —N(C₁-C₆ alkyl)₂. In some embodiments, R² is C₁-C₆alkyl, wherein each hydrogen atom in C₁-C₆ alkyl is independentlyoptionally substituted with one or more moieties selected from groupconsisting of —F, —OH, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), and—N(C₁-C₆ alkyl)₂, and R³ is H. In some embodiments, R² is methyl. Insome embodiments, R² is methyl, and R³ is H.

In some embodiments, R⁴ is H. In some embodiments, R⁴ is C₁-C₆ alkyl or3- to 7-membered heterocycloalkyl, wherein each hydrogen atom in C₁-C₆alkyl or 3- to 7-membered heterocycloalkyl is independently optionallysubstituted by halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl),—N(C₁-C₆ alkyl)₂, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆alkyl), —C(O)N(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or monocyclic 5- to7-membered heterocycloalkyl.

In some embodiments, R⁸ is C₁-C₆ alkyl, wherein each hydrogen atom isindependently optionally substituted by fluoro, chloro, bromo, —OH, —CN,—OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, C₃-C₇cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀ aryl, 5- to7-membered heteroaryl, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆alkyl), or —C(O)N(C₁-C₆ alkyl)₂. In some embodiments, R⁸ is ethyl,propyl, iso-propyl, or methylcyclopropyl.

In other embodiments, the compound of Formula I is selected from thegroup consisting of(7S)-3-amino-12-chloro-14-ethyl-11-fluoro-7-methyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo-[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one,(7S)-14-ethyl-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclo-tridecine-12-carbonitrile,14-ethyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile,(7S)-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile,(7S)-3-amino-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile,(7S)-3-amino-14-ethyl-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]-benzoxatriazacyclotridecine-12-carbonitrile,(7S)-3-amino-14-(cyclopropylmethyl)-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]-benzoxatriazacyclotridecine-12-carbonitrile,(7S)-3-amino-11-fluoro-7-methyl-4-oxo-14-(propan-2-yl)-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriaza-cyclotridecine-12-carbonitrile,or a pharmaceutically acceptable salt thereof.

The following represent illustrative embodiments of compounds of theFormula I:

Cpd Structure Name 1

(7S)-3-amino-12-chloro-14-ethyl-11-fluoro-7-methyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one 2

(7S)-14-ethyl-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 3

14-ethyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15- ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 4

(7S)-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 5

(7S)-3-amino-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 6

(7S)-3-amino-14-ethyl-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 7

(7S)-3-amino-14-(cyclopropylmethyl)-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15- ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 8

(7S)-3-amino-11-fluoro-7-methyl-4-oxo-14-(propan-2-yl)-4,5,6,7,13,14-hexahydro-1,15- ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 9

(7S)-3-amino-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 10

(7S)-3-amino-14-(²H₅)ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15- ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile 11

(7R)-3-amino-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12- carbonitrile

Those skilled in the art will recognize that the species listed orillustrated herein are not exhaustive, and that additional specieswithin the scope of these defined terms may also be selected.

Pharmaceutical Compositions

For treatment purposes, pharmaceutical compositions comprising thecompounds described herein may further comprise one or morepharmaceutically-acceptable excipients. A pharmaceutically-acceptableexcipient is a substance that is non-toxic and otherwise biologicallysuitable for administration to a subject. Such excipients facilitateadministration of the compounds described herein and are compatible withthe active ingredient. Examples of pharmaceutically-acceptableexcipients include stabilizers, lubricants, surfactants, diluents,anti-oxidants, binders, coloring agents, bulking agents, emulsifiers, ortaste-modifying agents. In preferred embodiments, pharmaceuticalcompositions according to the description are sterile compositions.Pharmaceutical compositions may be prepared using compounding techniquesknown or that become available to those skilled in the art.

Sterile compositions are also contemplated by the description, includingcompositions that are in accord with national and local regulationsgoverning such compositions.

The pharmaceutical compositions and compounds described herein may beformulated as solutions, emulsions, suspensions, or dispersions insuitable pharmaceutical solvents or carriers, or as pills, tablets,lozenges, suppositories, sachets, dragees, granules, powders, powdersfor reconstitution, or capsules along with solid carriers according toconventional methods known in the art for preparation of various dosageforms. Pharmaceutical compositions of the description may beadministered by a suitable route of delivery, such as oral, parenteral,rectal, nasal, topical, or ocular routes, or by inhalation. Preferably,the compositions are formulated for intravenous or oral administration.

For oral administration, the compounds the description may be providedin a solid form, such as a tablet or capsule, or as a solution,emulsion, or suspension. To prepare the oral compositions, the compoundsof the description may be formulated to yield a dosage of, e.g., fromabout 0.1 mg to 1 g daily, or about 1 mg to 50 mg daily, or about 50 to250 mg daily, or about 250 mg to 1 g daily. Oral tablets may include theactive ingredient(s) mixed with compatible pharmaceutically acceptableexcipients such as diluents, disintegrating agents, binding agents,lubricating agents, sweetening agents, flavoring agents, coloring agentsand preservative agents. Suitable inert fillers include sodium andcalcium carbonate, sodium and calcium phosphate, lactose, starch, sugar,glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol, andthe like. Exemplary liquid oral excipients include ethanol, glycerol,water, and the like. Starch, polyvinyl-pyrrolidone (PVP), sodium starchglycolate, microcrystalline cellulose, and alginic acid are exemplarydisintegrating agents. Binding agents may include starch and gelatin.The lubricating agent, if present, may be magnesium stearate, stearicacid, or talc. If desired, the tablets may be coated with a materialsuch as glyceryl monostearate or glyceryl distearate to delay absorptionin the gastrointestinal tract, or may be coated with an enteric coating.

Capsules for oral administration include hard and soft gelatin capsules.To prepare hard gelatin capsules, active ingredient(s) may be mixed witha solid, semi-solid, or liquid diluent. Soft gelatin capsules may beprepared by mixing the active ingredient with water, an oil, such aspeanut oil or olive oil, liquid paraffin, a mixture of mono anddi-glycerides of short chain fatty acids, polyethylene glycol 400, orpropylene glycol.

Liquids for oral administration may be in the form of suspensions,solutions, emulsions, or syrups, or may be lyophilized or presented as adry product for reconstitution with water or other suitable vehiclebefore use. Such liquid compositions may optionally contain:pharmaceutically-acceptable excipients such as suspending agents (forexample, sorbitol, methyl cellulose, sodium alginate, gelatin,hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel andthe like); non-aqueous vehicles, e.g., oil (for example, almond oil orfractionated coconut oil), propylene glycol, ethyl alcohol, or water;preservatives (for example, methyl or propyl p-hydroxybenzoate or sorbicacid); wetting agents such as lecithin; and, if desired, flavoring orcoloring agents.

For parenteral use, including intravenous, intramuscular,intraperitoneal, intranasal, or subcutaneous routes, the agents of thedescription may be provided in sterile aqueous solutions or suspensions,buffered to an appropriate pH and isotonicity or in parenterallyacceptable oil. Suitable aqueous vehicles include Ringer's solution andisotonic sodium chloride. Such forms may be presented in unit-dose formsuch as ampoules or disposable injection devices, in multi-dose formssuch as vials from which the appropriate dose may be withdrawn, or in asolid form or pre-concentrate that can be used to prepare an injectableformulation. Illustrative infusion doses range from about 1 to 1000m/kg/minute of agent admixed with a pharmaceutical carrier over a periodranging from several minutes to several days.

For nasal, inhaled, or oral administration, the inventive pharmaceuticalcompositions may be administered using, for example, a spray formulationalso containing a suitable carrier. The inventive compositions may beformulated for rectal administration as a suppository.

For topical applications, the compounds of the present description arepreferably formulated as creams or ointments or a similar vehiclesuitable for topical administration. For topical administration, theinventive compounds may be mixed with a pharmaceutical carrier at aconcentration of about 0.1% to about 10% of drug to vehicle. Anothermode of administering the agents of the description may utilize a patchformulation to affect transdermal delivery.

Methods of Treatment

As used herein, the terms “treat” or “treatment” encompass both“preventative” and “curative” treatment. “Preventative” treatment ismeant to indicate a postponement of development of a disease, a symptomof a disease, or medical condition, suppressing symptoms that mayappear, or reducing the risk of developing or recurrence of a disease orsymptom. “Curative” treatment includes reducing the severity of orsuppressing the worsening of an existing disease, symptom, or condition.Thus, treatment includes ameliorating or preventing the worsening ofexisting disease symptoms, preventing additional symptoms fromoccurring, ameliorating or preventing the underlying systemic causes ofsymptoms, inhibiting the disorder or disease, e.g., arresting thedevelopment of the disorder or disease, relieving the disorder ordisease, causing regression of the disorder or disease, relieving acondition caused by the disease or disorder, or stopping the symptoms ofthe disease or disorder.

The term “subject” refers to a mammalian patient in need of suchtreatment, such as a human. As used herein “cancer” includes any cancerknown in the art, particularly those cancers where SRC and MET, and/orCSF1R have been implicated in the disease. Examples of cancer typesinclude, but are not limited to, carcinomas, sarcomas, lymphomas,Hodgekin's disease, melanomas, mesotheliomas, Burkitt's lymphoma,nasopharyngeal carcinomas, leukemias, and myelomas. Examples of specificcancers include, but are not limited to, oral cancer, thyroid cancer,endocrine cancer, skin cancer, gastric cancer, esophageal cancer,laryngeal cancer, pancreatic cancer, colon cancer, bladder cancer, bonecancer, ovarian cancer, cervical cancer, uterine cancer, breast cancer,testicular cancer, prostate cancer, renal cancer, rectal cancer, kidneycancer, liver cancer, glioblastoma, or head & neck cancer, and lungcancers, such as non-small cell lung cancer, small cell lung cancer, andthe like.

In one aspect, the compounds and pharmaceutical compositions of thedescription specifically target SRC and MET, and/or CSF1R. Thus, thesecompounds and pharmaceutical compositions can be used to prevent,reverse, slow, or inhibit the activity of one or more of SRC and MET,and/or CSF1R. In some embodiments, methods of treatment of cancercomprising administering a therapeutically effective amount of acompound that inhibits one or more of SRC and MET, and/or CSF1R aredescribed herein. In other embodiments, methods for treating cancercomprising a. administering a therapeutically effective amount of acompound as described herein that inhibits one or more of SRC and MET,and/or CSF1R are described. In other embodiments, methods for treatingcancer comprising a. administering a therapeutically effective amount ofa compound as described herein are described. In other embodiments, thecancer is gastric cancer, colon cancer, renal cancer, liver cancer, lungcancer, glioblastoma, or head & neck cancer.

In some embodiments, the disclosure is directed to a compound thatinhibits SRC and MET, and/or CSF1R, or a pharmaceutically acceptablesalt thereof, for use in the treatment of cancer in a patient. In someembodiments, the compound that inhibits SRC and MET, and/or CSF1R is ofthe Formula I. In some embodiments, the cancer is gastric cancer, coloncancer, renal cancer, liver cancer, lung cancer, glioblastoma, or head &neck cancer.

In some embodiments, the disclosure is directed to use of a compoundthat inhibits SRC and MET, and/or CSF1R, or a pharmaceuticallyacceptable salt thereof, for use in the treatment of cancer in apatient. In some embodiments, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments, the cancer isgastric cancer, colon cancer, renal cancer, liver cancer, lung cancer,glioblastoma, or head & neck cancer.

In some embodiments, the disclosure is directed to use of a compoundthat inhibits SRC and MET, and/or CSF1R, or a pharmaceuticallyacceptable salt thereof, in the preparation of a medicament for thetreatment of cancer in a patient. In some embodiments, the compound thatinhibits SRC and MET, and/or CSF1R is of the Formula I. In someembodiments, the cancer is gastric cancer, colon cancer, renal cancer,liver cancer, lung cancer, glioblastoma, or head & neck cancer.

In some embodiments, the disclosure is directed to a compositioncomprising a compound that inhibits SRC and MET, and/or CSF1R, or apharmaceutically acceptable salt thereof, in a therapeutically effectiveamount, for use in the treatment of cancer in a patient. In someembodiments, the compound that inhibits SRC and MET, and/or CSF1R is ofthe Formula I. In some embodiments, the cancer is gastric cancer, coloncancer, renal cancer, liver cancer, lung cancer, glioblastoma, or head &neck cancer.

In the inhibitory methods of the description, an “effective amount”means an amount sufficient to inhibit the target. Measuring such targetmodulation may be performed by routine analytical methods such as thosedescribed below. Such modulation is useful in a variety of settings,including in vitro assays. In such methods, the cell can be a cancercell with abnormal signaling due to upregulation, mutation, aberrantactivity of, and/or changes in SRC and MET, and/or CSF1R.

In treatment methods according to the description, an “effective amount”means an amount or dose sufficient to generally bring about the desiredtherapeutic benefit in subjects needing such treatment. Effectiveamounts or doses of the compounds of the description may be ascertainedby routine methods, such as modeling, dose escalation, or clinicaltrials, taking into account routine factors, e.g., the mode or route ofadministration or drug delivery, the pharmacokinetics of the agent, theseverity and course of the infection, the subject's health status,condition, and weight, and the judgment of the treating physician. Anexemplary dose is in the range of about from about 0.1 mg to 1 g daily,or about 1 mg to 50 mg daily, or about 50 to 250 mg daily, or about 250mg to 1 g daily. The total dosage may be given in single or divideddosage units (e.g., BID, TID, QID).

Once improvement of the patient's disease has occurred, the dose may beadjusted for preventative or maintenance treatment. For example, thedosage or the frequency of administration, or both, may be reduced as afunction of the symptoms, to a level at which the desired therapeutic orprophylactic effect is maintained. Of course, if symptoms have beenalleviated to an appropriate level, treatment may cease. Patients may,however, require intermittent treatment on a long-term basis upon anyrecurrence of symptoms. Patients may also require chronic treatment on along-term basis.

Drug Combinations

The inventive compounds described herein may be used in pharmaceuticalcompositions or methods in combination with one or more additionalactive ingredients in the treatment of the diseases and disordersdescribed herein. Further additional active ingredients include othertherapeutics or agents that mitigate adverse effects of therapies forthe intended disease targets. Such combinations may serve to increaseefficacy, ameliorate other disease symptoms, decrease one or more sideeffects, or decrease the required dose of an inventive compound. Theadditional active ingredients may be administered in a separatepharmaceutical composition from a compound of the present description ormay be included with a compound of the present description in a singlepharmaceutical composition. The additional active ingredients may beadministered simultaneously with, prior to, or after administration of acompound of the present description.

Combination agents include additional active ingredients are those thatare known or discovered to be effective in treating the diseases anddisorders described herein, including those active against anothertarget associated with the disease. For example, compositions andformulations of the description, as well as methods of treatment, canfurther comprise other drugs or pharmaceuticals, e.g., other activeagents useful for treating or palliative for the target diseases orrelated symptoms or conditions. Such additional agents include, but arenot limited to, kinase inhibitors, such as EGFR inhibitors (e.g.,erlotinib, gefitinib), Raf inhibitors (e.g., vemurafenib), VEGFRinhibitors (e.g., sunitinib), ALK inhibitors (e.g., crizotinib) standardchemotherapy agents such as alkylating agents, antimetabolites,anti-tumor antibiotics, topoisomerase inhibitors, platinum drugs,mitotic inhibitors, antibodies, hormone therapies, or corticosteroids.For pain indications, suitable combination agents includeanti-inflammatories such as NSAIDs. The pharmaceutical compositions ofthe description may additional comprise one or more of such activeagents, and methods of treatment may additionally comprise administeringan effective amount of one or more of such active agents.

In some embodiments, the disclosure is directed to a method of treatingcancer in a patient comprising, a. administering a therapeuticallyeffective amount of a compound that inhibits SRC and MET, and/or CSF1R;and b. administering a therapeutically effective amount of at least oneadditional anti-cancer agent. In some embodiments, the at least oneadditional anti-cancer agent is an EGFR inhibitor, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the additional anti-canceragent is an antibody of EGFR. In some embodiments, the compound thatinhibits SRC and MET, and/or CSF1R is of the Formula I. In someembodiments, the cancer is gastric cancer, liver cancer, lung cancer, orhead & neck cancer.

In some embodiments, the disclosure is directed to a compound thatinhibits SRC and MET, and/or CSF1R, or a pharmaceutically acceptablesalt thereof, in combination with a therapeutically effective amount ofat least one additional anti-cancer agent, or a pharmaceuticallyacceptable salt thereof, for use in the treatment of cancer in apatient. In some embodiments, the at least one additional anti-canceragent is an EGFR inhibitor, or a pharmaceutically acceptable saltthereof. In some embodiments, the additional anti-cancer agent is anantibody of EGFR. In some embodiments, the compound that inhibits SRCand MET, and/or CSF1R is of the Formula I. In some embodiments, thecancer is gastric cancer, liver cancer, lung cancer, or head & neckcancer.

In some embodiments, the disclosure is directed to use of a compoundthat inhibits SRC and MET, and/or CSF1R, or a pharmaceuticallyacceptable salt thereof, in combination with a therapeutically effectiveamount of at least one additional anti-cancer agent for the treatment ofcancer in a patient. In some embodiments, the compound that inhibits SRCand MET, and/or CSF1R is of the Formula I. In some embodiments of thisaspect, the cancer is gastric cancer, liver cancer, lung cancer, or head& neck cancer. In some embodiments, the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof. In some embodiments, the additional anti-cancer agent isan antibody of EGFR.

In some embodiments, the disclosure is directed to use of a compoundthat inhibits SRC and MET, and/or CSF1R, or a pharmaceuticallyacceptable salt thereof, in the preparation of a medicament for thetreatment of cancer in a patient in combination with a therapeuticallyeffective amount of at least one additional anti-cancer agent. In someembodiments, the compound that inhibits SRC and MET, and/or CSF1R is ofthe Formula I. In some embodiments of this aspect, the cancer is gastriccancer, liver cancer, lung cancer, or head & neck cancer. In someembodiments, the at least one additional anti-cancer agent is an EGFRinhibitor, or a pharmaceutically acceptable salt thereof. In someembodiments, the additional anti-cancer agent is an antibody of EGFR.

In some embodiments, the disclosure is directed to a compositioncomprising a compound that inhibits SRC and MET, and/or CSF1R, or apharmaceutically acceptable salt thereof, in a therapeutically effectiveamount, for use in the treatment of cancer in a patient. In someembodiments of this aspect, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments, the cancer isgastric cancer, liver cancer, lung cancer, or head & neck cancer. Insome embodiments, the compound is administered in combination with atherapeutically effective amount of at least one additional anti-canceragent. In some embodiments, the at least one additional anti-canceragent is an EGFR inhibitor, or a pharmaceutically acceptable saltthereof. In some embodiments, the additional anti-cancer agent is anantibody of EGFR.

In some embodiments, the disclosure relates to a synergistic compositionof a compound that inhibits SRC and MET, and/or CSF1R, and an EGFRinhibitor, where the two components come into contact with each other ata locus. In some embodiments, the compound that inhibits SRC and MET,and/or CSF1R is of the Formula I. In some embodiments, the at least oneadditional anti-cancer agent is an EGFR inhibitor, or a pharmaceuticallyacceptable salt thereof. In some embodiments, the additional anti-canceragent is an antibody of EGFR.

EXAMPLES Chemical Synthesis

Exemplary chemical entities useful in methods of the description willnow be described by reference to illustrative synthetic schemes fortheir general preparation below and the specific examples that follow.Artisans will recognize that, to obtain the various compounds herein,starting materials may be suitably selected so that the ultimatelydesired substituents will be carried through the reaction scheme with orwithout protection as appropriate to yield the desired product.Alternatively, it may be necessary or desirable to employ, in the placeof the ultimately desired substituent, a suitable group that may becarried through the reaction scheme and replaced as appropriate with thedesired substituent. Furthermore, one of skill in the art will recognizethat the transformations shown in the schemes below may be performed inany order that is compatible with the functionality of the particularpendant groups.

Abbreviations

The examples described herein use materials, including but not limitedto, those described by the following abbreviations known to thoseskilled in the art:

g grams eq equivalents mmol millimoles mL milliliters EtOAc ethylacetate MHz megahertz ppm parts per million δ chemical shift s singlet ddoublet t triplet q quartet quin quintet br broad m multiplet Hz hertzTHF tetrahydrofuran ° C. degrees Celsius PE petroleum ether EA ethylacetate R_(f) retardation factor N normal J coupling constant DMSO-d₆deuterated dimethyl sulfoxide n-BuOH n-butanol DIEAn.n-diisopropylethylamine TMSCl trimethylsilyl chloride min minutes hrhours Me methyl Et ethyl i-Pr isopropyl TLC thin layer chromatography Mmolar Compd# compound number MS mass spectrum m/z mass-to-charge ratioMs methanesulfonyl FDPP pentafluorophenyl diphenylphosphinate Boctert-butyloxycarbonyl TFA trifluoroacetic acid Tos toluenesulfonyl DMAP4-(dimethylamino)pyridine μM micromolar ATP adenosine triphosphate IC₅₀half maximal inhibitory concentration U/mL units of activity permilliliter KHMDS potassium bis(trimethylsilyl)amide DIAD diisopropylazodicarboxylate MeTHF 2-methyltetrahydrofuran MOM methoxymethyl DCMdichloromethane DMF N,N-dimethyl formamide DPPA diphenyl phosphorylazide DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DIPEAN,N-diisopropylethylamine

General Method A.

Preparation 2-chloro-3-fluoro-6-hydroxybenzaldehyde (A-1-4)

Step 1. To a solution of A-1-1 (20.00 g, 136.47 mmol, 1.00 eq.) andsodium hydride (6.55 g, 60% purity, 272.94 mmol, 2.00 eq.) in DMF(200.00 mL) was added MOMCl (21.97 g, 272.94 mmol, 20.73 mL, 2.00 eq.)at 0° C. under N₂. The mixture was stirred at 25° C. for 10 hours. TLC(Petroleum ether/Ethyl acetate=5/1) showed the starting material wasconsumed completely and one new spot was found. The reaction mixture wasquenched by water (150 mL), and then diluted with water (150 mL) andextracted with ethyl acetate (3×100 mL). The combined organic layerswere washed with brine (150 mL), dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure to give A-1-2 (20.00 g,76.89% yield) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ: 7.11 (dd,J=2.8, 6.0 Hz, 1H), 7.04 (t, J=8.8 Hz, 1H), 6.90 (td, J=3.2, 9.2 Hz,1H), 5.12 (s, 2H), 3.47 (s, 3H).

Step 2. To a solution of A-1-2 (20.00 g, 104.93 mmol, 1.00 eq.) in THF(250.00 mL) was added n-BuLi (2.5 M, 125.92 mL, 3.00 eq.) at −65° C.under N₂. The mixture was stirred at −65° C. for 2 hours. The mixturewas quenched by DMF (76.69 g, 1.05 mol, 80.73 mL, 10.00 eq.) and themixture was stirred at −65° C. for 15 min under N₂. TLC (Petroleumether: Ethyl acetate=3:1) showed the starting material was consumedcompletely and one new spot was found. The reaction mixture was dilutedwith water (300 mL) and extracted with ethyl acetate (150 mL*3). Thencombined organic layers and dried over anhydrous sodium sulfate,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂, Petroleumether/Ethyl acetate=I/O to 1/1) to give A-1-3 (4.80 g, 20.93% yield) asa colorless oil. ¹H NMR (400 MHz, CDCl₃) δ: 10.48 (s, 1H), 7.28 (t,J=8.8 Hz, 1H), 7.15 (dd, J=4.0, 9.2 Hz, 1H), 5.25 (s, 2H), 3.51 (s, 3H).

Step 3. To a solution of A-1-3 (4.00 g, 18.3 mmol, 1.00 eq.) inHCl/dioxane (40.0 mL) was stirred at 25° C. for 2 hours. The reactionmixture was diluted with water (30.0 mL) and extracted with ethylacetate (25.0 mL×3). The combined organic layers were washed with water(30.0 mL), dried over anhydrous sodium sulfate, filtered andconcentrated under reduced pressure to give A-1-4 (2.50 g, 14.3 mmol,yield=78.3%) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ=11.68 (s, 1H),10.43 (s, 1H), 7.37-7.32 (m, 1H), 6.91 (dd, J=4.0, 9.2 Hz, 1H).

General Method B.

Preparation of 2-bromo-3-fluoro-6-hydroxybenzaldehyde (A-2-4)

Step 1. To a solution of A-2-1 (30.0 g, 155 mmol, 1 eq.) in THF (300 mL)was added LDA (2 M, 116 mL, 1.5 eq.) at −78° C. and stirred for 1 hour,then DMF (34.1 g, 466 mmol, 3 eq.) was added at −78° C. and stirred for2 hours. The reaction mixture was quenched by addition saturatedammonium chloride (200 mL) at 0° C., then diluted with water (300 mL)and extracted with ethyl acetate (1.00 L). The organic layer was washedby brine (200 mL), dried over anhydrous sodium sulfate, filtered andconcentrated. The crude product was purified by column chromatography togive A-2-2 (20.0 g, 90.5 mmol, yield=58.2%) as yellow solid. ¹H NMR (400MHz, CDCl₃) δ=10.34 (s, 1H), 7.37-7.34 (m, 1H), 7.17-7.34 (m, 1H).

Step 2. A solution of A-2-2 (20.0 g, 90.5 mmol, 1.00 eq.) in THF (100mL) and methanol (240 mL) was heated to 60° C., then a solution ofsodium methylate (4.3 M, 25.3 mL, 1.2 eq.) in methanol was added andstirred at 60° C. for 12 hours. The reaction mixture was quenched byaddition water (200 mL) and extracted with ethyl acetate (500 mL). Theorganic layer was washed by brine (100 mL), dried over anhydrous sodiumsulfate, filtered and concentrated, and the residue was purified bycolumn chromatography to give A-2-3 (13.5 g, 57.9 mmol, yield=64.0%) asa yellow solid. ¹H NMR (400 MHz, CDCl₃) δ=10.30 (s, 1H), 7.20 (dd,J=7.6, 9.2 Hz, 1H), 6.86 (dd, J=4.0, 9.2 Hz, 1H), 3.84 (s, 3H).

Step 3. To a solution of A-2-3 (13.0 g, 55.8 mmol, 1.00 eq.) in DCM (150mL) was added BBr₃ (28.0 g, 112 mmol, 2.00 eq.) at −40° C. drop-wise,then the mixture was stirred at 0° C. for 3 hours. The reaction mixturewas quenched by addition methanol (20.0 mL) and saturated sodiumbicarbonate solution (50.0 mL) at 0° C., then extracted with ethylacetate (300 mL). The organic layer was washed by brine (100 mL), driedover anhydrous sodium sulfate, filtered and concentrated, and theresidue was purified by column chromatography to give A-2-4 (10.5 g,43.2 mmol, yield=77.4%) as a yellow solid. ¹HNMR (400 MHz, CDCl₃)δ=11.78 (s, 1H), 10.35 (s, 1H), 7.32 (dd, J=7.6, 9.2 Hz, 1H), 6.96 (dd,J=4.0, 9.2 Hz, 1H).

General Method C.

Preparation of ethyl2-amino-5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (A-6-7)

Step 1. To a solution of A-3-1 (100 g, 884 mmol, 1.00 eq.) and A-3-1A(230 g, 1.59 mol, 1.80 eq.) in ethanol (1.50 L) was added TEA (4.47 g,44.2 mmol, 0.05 eq.) at 0° C. The mixture was stirred at 25° C. for 12hours. The solvent was removed to give the crude product which waspurified by column chromatography to give A-3-2 (200 g, 738 mmol,yield=83.5%) as off-white oil. ¹H NMR (400 MHz, CDCl₃) δ=10.21 (br s,1H), 6.95 (br s, 1H), 4.29-4.34 (m, 2H), 1.37 (t, J=7.2 Hz, 3H).

Step 2. To a solution of ethyl A-3-2 (100 g, 388 mmol, 1.00 eq.) in DMF(500 mL) was added hydrazine hydrate (311 g, 3.11 mol, 50.0% purity,8.00 eq.). The mixture was stirred at 100° C. for 2 hours. Removed thesolvent and added DCM (500 mL), the resulting mixture was stirred for 12hours. The solid was filtered and washed with DCM (200 mL) to give A-3-3(60.0 g, 317 mmol, yield=81.7%) as brown solid. ¹H NMR (400 MHz,DMSO-d₆) δ=7.91 (s, 1H), 7.50 (s, 2H), 4.57 (s, 2H), 4.03 (q, J=7.2 Hz,2H), 1.16 (t, J=7.2 Hz, 3H).

Step 3. To a solution of fresh prepared sodium ethoxide (0.50 M, 2.35 L,4.00 eq.) in ethanol (200 mL) was added A-3-3 (50.0 g, 294 mmol, 1.00eq.), then A-3-3A (41.2 g, 294 mmol, 1.00 eq.) was added. The mixturewas stirred at 90° C. for 9 hours. Filtered and filter cake was washedwith ethanol (100 mL) to give the A-3-4 (25.0 g, 113 mmol, yield=38.3%)as a brown solid. ¹H NMR (400 MHz, DMSO-d₆) δ=7.71 (d, J=7.2 Hz, 1H),5.57-5.46 (m, 3H), 4.15 (q, J=7.2 Hz, 2H), 1.25 (t, J=7.2 Hz, 3H).

Step 4. To a solution of A-3-4 (18.0 g, 81.0 mmol, 1.00 eq.) in DCM (300mL) was added triethylamine (20.5 g, 202 mmol, 2.50 eq.) at 0° C., thentrifluoroacetic anhydride (20.4 g, 97.2 mmol, 1.20 eq.) was added. Themixture was stirred at 25° C. for 12 hours. The solid was collected byfiltration and washed with DCM (100 mL) to give A-3-5 (18.0 g, 47.4mmol, yield=58.5%) as a yellow solid. LCMS:EW6129-170-P1D (M+1:319.1).

Step 5. A solution of A-3-5 (18.0 g, 56.6 mmol, 1.00 eq.) in freshdistilled POCl₃ (180 mL) was stirred at 100° C. for 5 hours. The mixturewas poured into ice-water (500 mL) at 0° C., filtered and filter cakewashed with water (200 mL) and then collected to give A-3-6 (15.0 g,43.6 mmol, yield=77.1%) as a black brown solid. ¹H NMR (400 MHz,DMSO-d₆) δ=11.93 (s, 1H), 9.31 (d, J=7.2 Hz, 1H), 7.47 (d, J=7.2 Hz,1H), 4.28 (q, J=7.2 Hz, 2H), 1.28 (br t, J=7.2 Hz, 3H).

Step 6. To a solution of A-3-6 (13.0 g, 38.6 mmol, 1.00 eq.) inn-butanol (150 mL) and acetonitrile (150 mL) was added potassiumcarbonate (10.7 g, 77.2 mmol, 2.00 eq.). The mixture was stirred at 60°C. for 8 hours. The reaction mixture was quenched by addition water (200mL) and extracted with dichloromethane/methanol=10/1 (500 mL×3). Thecombined organic layers were washed with brine (300 mL), dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure to give a residue. The residue was purified by Prep-HPLC (basiccondition) to give the A-3-7 (4.8 g, 19.2 mmol, yield=49.6%) as whitesolid. ¹H NMR (400 MHz, CDCl₃) δ=8.23 (d, J=7.2 Hz, 1H), 6.74 (d, J=7.2Hz, 1H), 5.44 (s, 2H), 4.37 (q, J=7.2 Hz, 2H), 1.37 (t, J=7.2 Hz, 3H).

General Method D.

Preparation of ethyl2-amino-5-((2-chloro-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-1)

Step 1. A solution of A-1-4 (166 mg, 951 mol, 1 eq.) and ethylamine (129mg, 2.85 mmol, 3.0 eq.) in methanol (4.8 mL) was stirred for 1 hour at65° C. The reaction mixture was cooled to room temperature and NaBH₄ (53mg, 1.4 mmol, 1.5 eq.) was added, the reaction mixture was stirred at25° C. for 30 min. The mixture was quenched with water (15 mL) andstirred for 5 min. The mixture was extracted with DCM (3×15 mL), driedwith Na₂SO₄ and concentrated under reduced pressure. Flashchromatography (ISCO system, silica (12 g), 0-100% ethyl acetate inhexane) provide C₉H₁₁OFClN (175.3 mg, 860.8 mol, 90.5% yield).

Step 2. To a mixture of A-4-1 (97.3 mg, 0.477 mmol, 1.15 eq.) and A-3-7(100 mg, 0.415 mmol, 1.0 eq.) in n-butanol (2.00 mL) was added DIEA (269mg, 2.1 mmol, 5.00 eq.). The mixture was heated to 85° C. and stirredfor 20 hours. Removed the solvent and the residue was purified by columnchromatography to give compound A-1 (146 mg, 357 mol, yield=86%).

General Method E.

Preparation of ethyl5-((2-cyano-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-2)

Step 1. To a solution of A-5-1 (2.00 g, 9.95 mmol, 1.00 eq.) in DMF(20.00 mL) was added sodium hydride (796 mg, 19.9 mmol, 60% purity, 2.00eq.) at 0° C. under N₂ atmosphere. The mixture was stirred at 0° C. for30 mins, then chloro(methoxy)methane (1.20 g, 14.92 mmol, 1.13 mL, 1.50eq.) was added at 0° C. The mixture was stirred at 20° C. for 3 hours.Then the mixture was quenched by water (100 mL) and extracted with ethylacetate (50.0 mL×3). The organic layer washed by brine (100 mL) anddried over anhydrous sodium sulfate, filtered and concentrated to giveA-5-2 (2.70 g, crude) as a yellow solid. LCMS:EW6129-85-P1A(M+23:268.9).

Step 2. To a solution of A-5-2 (2.70 g, 11.0 mmol, 1.00 eq.) andethanamine (745 mg, 16.5 mmol, 1.08 mL, 1.50 eq.) in methanol (20.0 mL)was added sodium acetate (1.08 g, 13.2 mmol, 1.20 eq.) in one portion at20° C. under N₂ atmosphere. The mixture was stirred at 20° C. for 30mins, then sodium cyanoborohydride (1.04 g, 16.5 mmol, 1.50 eq.) wasadded and stirred at 20° C. for 15 hours. The mixture was concentrated,diluted with water (30.0 mL) and extracted with ethyl acetate (15.0mL×3). The combined organic layer washed by brine (30.0 mL), dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure to give A-5-3 (2.95 g, 10.7 mmol, yield=97.6%) as a yellowsolid. LCMS:EW6129-100-P1B(M+1:274).

Step 3. To a solution of A-5-3 (2.95 g, 10.7 mmol, 1.00 eq.) and A-5-3A(2.43 g, 10.7 mmol, 1.00 eq.) in n-BuOH (20.0 mL) was added DIEA (5.56g, 43.0 mmol, 7.51 mL, 4.00 eq.) in one portion at 20° C. under N₂atmosphere. The mixture was heated to 95° C. and stirred for 2 hours.Then the mixture was diluted with water (50.0 mL) and extracted withethyl acetate (30.0 mL×3). The organic layer washed by brine (50.0 mL)and dried over anhydrous sodium sulfate. The residue was purified bycolumn chromatography (SiO₂, Petroleum ether/ethyl acetate=5/1 to 1:1)to give A-5-4 (1.66 g, 3.58 mol, yield=33.3%) as a yellow solid. ¹H NMR(400 MHz, CDCl₃) δ=8.32-8.28 (m, 2H), 7.30-7.28 (m, 1H), 7.17 (t, J=8.4Hz, 1H), 7.14-7.08 (m, 1H), 6.55 (br s, 1H), 5.32-5.15 (m, 2H), 5.11 (s,2H), 4.34 (q, J=7.2 Hz, 2H), 3.51 (s, 2H), 3.34 (s, 3H), 1.38 (t, J=7.2Hz, 3H), 1.14 (t, J=7.2 Hz, 3H).

Step 4. To a solution of A-5-4 (1.50 g, 3.24 mmol, 1.00 eq.) in DMF(20.0 mL) was added Pd(dppf)Cl₂ (237 mg, 324 mol, 0.10 eq.), Zn(CN)₂(570 mg, 4.86 mmol, 308 μL, 1.50 eq.) and Zn (10.6 mg, 162 mol, 0.05eq.) at 20° C. under N₂ atmosphere. The mixture was heated to 120° C.and stirred for 15 hours. The mixture was diluted with water (100 mL)and extracted with ethyl acetate (50.0 mL×3). The organic layer wascombined and washed by brine (100 mL) and dried over anhydrous sodiumsulfate. The residue was purified by column chromatography (SiO₂,Petroleum ether/ethyl acetate=10/1 to 1:1) to give A-5-5 (830 mg, 2.03mmol, yield=62.7%) as a yellow oil. LCMS:EW6129-107-P1A(M+1:410.2).

Step 5. To a solution of A-5-5 (730 mg, 1.78 mmol, 1.00 eq.) inHCl/dioxane (30.0 mL) was stirred at 20° C. for 3 hours. The reactionmixture was concentrated under reduced pressure to give A-2 (630 mg,1.72 mmol, yield=96.6%) as a white solid. ¹H NMR (400 MHz, CDCl₃)δ=10.84 (br s, 1H), 8.37 (d, J=7.6 Hz, 1H), 8.34 (s, 1H), 7.36-7.31 (m,1H), 7.24-7.20 (m, 2H), 6.44 (br d, J=7.6 Hz, 1H), 5.14 (s, 2H), 4.43(q, J=7.2 Hz, 2H), 3.72-3.67 (m, 2H), 1.40 (t, J=7.2 Hz, 3H), 1.34 (t,J=7.2 Hz, 3H).

General Method F

Preparation of ethyl5-((2-bromo-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-3)

Step 1. A solution of A-2-4 (3.00 g, 13.7 mmol, 1 eq.) and ethanamine(1.24 g, 27.4 mmol, 2.00 eq.) in methanol (30.0 mL) was stirred for 30min at 25° C. and then NaBH₄ (1.04 g, 27.4 mmol, 2.00 eq.) was added,the reaction mixture was stirred at 25° C. for 12 hours. The solvent wasremoved and the result mixture was diluted with water (20 mL), extractedwith ethyl acetate (100 mL). The organic layer was washed by brine (100mL), dried over anhydrous sodium sulfate, filtered and concentrated togive A-6-1 (2.40 g, 8.71 mmol, yield=63.6%) as a white solid. ¹HNMR (400MHz, CDCl₃) δ=6.93 (t, J=8.4 Hz, 1H), 6.71 (dd, J=4.4, 8.4 Hz, 1H), 4.23(s, 2H), 2.76 (q, J=7.2 Hz, 2H), 1.19 (t, J=7.2 Hz, 3H).

Step 2. To a mixture of A-6-1 (1.20 g, 4.84 mmol, 1.00 eq.) and A-5-3A(1.31 g, 5.80 mmol, 1.20 eq.) in n-butanol (10.0 mL) was added DIEA(2.50 g, 19.4 mmol, 4.00 eq.) in one portion at 25° C. under N₂protecting. The mixture was heated to 95° C. and stirred for 2 hours.Removed the solvent and the residue was purified by columnchromatography to give compound A-3 (1.20 g, 2.37 mmol, yield=49.0%) asa white solid. ¹HNMR (400 MHz, CDCl₃) δ=10.40 (s, 1H), 8.37-8.29 (m,2H), 7.03 (dd, J=8.0, 8.8 Hz, 1H), 6.88 (dd, J=4.8, 8.8 Hz, 1H), 6.40(d, J=8.0 Hz, 1H), 5.18 (br s, 2H), 4.40 (q, J=7.2 Hz, 2H), 3.65 (q,J=7.2 Hz, 2H), 1.38 (t, J=7.2 Hz, 3H), 1.31 (t, J=7.2 Hz, 3H).

General Method G.

Preparation of ethyl2-amino-5-((2-bromo-3-fluoro-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-4)

Step 1. To a mixture of A-6-1 (0.30 g, 1.21 mmol, 1.00 eq.) and A-3-7(349 mg, 1.45 mmol, 1.2 eq.) in n-butanol (5.00 mL) was added DIEA (625mg, 4.84 mmol, 4.00 eq.) in one portion at 25° C. under N₂ protecting.The mixture was heated to 95° C. and stirred for 2 hours. Removed thesolvent and the residue was purified by column chromatography to givecompound A-4 (250 mg, 514 mol, yield=42.5%) as a yellow solid. ¹HNMR(400 MHz, CDCl₃) δ=(br s, 1H), 8.05 (d, J=7.6 Hz, 1H), 7.04 (dd, J=8.0,8.8 Hz, 1H), 6.85 (dd, J=4.8, 8.8 Hz, 1H), 6.18 (d, J=7.8 Hz, 1H), 5.29(s, 2H), 5.16 (br s, 2H), 4.40 (q, J=7.2 Hz, 2H), 3.58 (q, J=7.2 Hz,2H), 1.39 (t, J=7.2 Hz, 3H), 1.29 (t, J=7.2 Hz, 3H).

Preparation of ethyl2-amino-5-((2-bromo-6-hydroxybenzyl)(ethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-5)

General method F and G were used to make A-5 starting with A-5-1 ingeneral method E.

Preparation of Ethyl2-amino-5-((2-bromo-3-fluoro-6-hydroxybenzyl)(cyclopropylmethyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-6)

General method D was used to make A-6.

Preparation of ethyl2-amino-5-((2-bromo-3-fluoro-6-hydroxybenzyl)(isopropyl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-7)

General method D was used to make A-7.

General Method H.

Preparation of ethyl2-amino-5-{[(2-bromo-3-fluoro-6-hydroxyphenyl)methyl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-8)

Step 1. To a solution of A-2-4 (250 mg, 1.14 mmol) andchloro(methoxy)methane (119 mg, 1.48 mmol, 113 μL) in THF (5.7 mL) wasadded DIEA (368 mg, 2.85 mmol) at −78° C. under Ar atmosphere. Themixture was slowly warmed to 25° C. and stirred for 14 hours. Then themixture was quenched by water (10 mL) and extracted with DCM (3×15 mL).The organic layer was dried over anhydrous sodium sulfate, filtered andconcentrated. Flash chromatography (ISCO system, silica (12 g), 5-15%ethyl acetate in hexane) provided A-8-1 (96.6 mg, 32% yield).

Step 2. To a solution of A-8-1 (96.6 mg, 0.367 mmol) and A-8-1A (111 mg,0.918 mmol) in THF (1.0 mL), Me-THF (1.0 mL) and diglyme (52 μL) wasadded Ti(OEt)₄ (586 mg, 2.57 mmol, 538 μL) under Ar atmosphere. Themixture was heated to 75° C. and stirred for 2 hours. The mixture wascooled to room temperature and poured into a 5:1 MeOH:water solution (60mL). To this suspension was added celite and the mixture filteredthrough a bed a celite. The celite pad was washed with MeOH (50 mL) andethyl acetate (50 mL). Combined filtrates were added to water (100 mL)and the mixture extracted with ethyl acetate (3×75 mL). The combinedorganic extracts were washed with brine (50.0 mL), dried over anhydroussodium sulfate, filtered and concentrated under reduced pressure. Flashchromatography (ISCO system, silica (12 g), 0-30% ethyl acetate inhexane) provided A-8-2 (101.6 mg, 75% yield).

Step 3. To a solution of A-8-2 (101.6 mg, 0.28 mmol) and water (15.0 mg,0.83 mmol) in THF (1.4 mL) at −78° C. was added NaBH₄ (31.5 mg, 0.83mmol) in one portion. The mixture was slowly warmed to 25° C. andstirred for 14 hours. Then the mixture was cooled to −20° C. andquenched with water (10.0 mL) and extracted with DCM (3×15 mL). Combinedextracts were dried with Na₂SO₄ then concentrated under reducedpressure. Flash chromatography (ISCO system, silica (12 g), 30-60% ethylacetate in hexane) provided A-8-3 (quantitative).

Step 4. To a solution of A-8-3 (102 mg, 0.28 mmol, 1.00 eq.) in DCM (4.0mL) was added 4M HCl in dioxane (3.0 mL). The reaction mixture wasstirred at 25° C. for 1.5 hours then concentrated under reducedpressure. The solids were suspended in DCM (5 mL) and saturatedbicarbonate solution (5 mL) was added and the mixture stirred for 5 min.The mixture was extracted with DCM (3×15 mL). Combined extracts weredried with Na₂SO₄ then concentrated under reduced pressure. Flashchromatography (ISCO system, silica (4 g), 80-100% ethyl acetate inhexane) provided A-8-4 (53.5 mg, 88% yield).

Step 5. General method G was used to make A-8 starting with A-8-4.

MS Compd# Structure m/z A-1

408.1 A-2

366.1 A-3

437.0 A-4

452.3 A-5

434.2 A-6

478.3 A-7

466.0 A-8

424.0

General Method H.

Preparation of(7S)-3-amino-12-chloro-14-ethyl-11-fluoro-7-methyl-6,7,13,14-tetrahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecin-4(5H)-one(1)

Step 1. To a solution of azeotrope dried phenol A-1 (50 mg, 0.12 mmol)and (R)-tert-butyl (2-hydroxypropyl)carbamate (25.8 mg, 0.147 mmol) indichloromethane (300 μL) was added PPh3 (40.2 g, 0.153 mmol). Themixture was stirred until completely dissolved then cooled to 0° C. andDIAD (32.2 mg, 0.159 mmol, 31.3 μL) was added dropwise with mixing. Themixture was warmed to 35° C. and stirred for 1 hour. Flashchromatography (ISCO system, silica (12 g), 0-100% ethyl acetate inhexane) provided impure 1-1.

Step 2. To a solution of 1-7 (69.2 mg, 122 μmol) in MeOH (4 mL) and THF(2 mL) at ambient temperature was added aqueous LiOH solution (2.0 M, 2mL). The mixture was heated at 70° C. for 25 hours, cooled to 20° C.then quenched with aqueous HCl solution (2.0 M) to acidic. The mixturewas extracted with DCM (3×5 mL), dried with Na₂SO₄, concentrated underreduced pressure, and dried under high vacuum. The crude material wasdissolved in DCM (4 mL) followed by addition of HCl in 1,4-dioxane (4 M,3 mL). The mixture was stirred ambient temperature for 1 hour,concentrated under reduced pressure, and dried under high vacuum. Thecrude material was dissolved in in DMF (2.0 mL) and DCM (8.0 mL) andHünig's base (158 mg, 1.22 mmol, 213 μL) then FDPP (61.2 mg, 159 μmol)was added in one portion. The reaction was stirred for 3 hours thenquenched with 2 M Na₂CO₃ solution (5 mL). Mixture was stirred for 5 minthen extracted with DCM (4×10 mL). Combined extracts were dried withNa₂SO₄ and concentrated under reduced pressure. Flash chromatography(ISCO system, silica (12 g), 0-7.5% methanol in dichloromethane)provided 1 (11.1 mg, 26.5 μmol, 21% yield).

General Method I.

Preparation of(7S)-14-ethyl-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile(2)

Step 1. To a solution of azeotrope dried phenol A-2 (100 mg, 0.274 mmol)and (R)-tert-butyl (2-hydroxypropyl)carbamate (95.9 mg, 0.547 mmol) indichloromethane (182 μL) was added PPh3 (144 mg, 0.547 mmol). Themixture was stirred until completely dissolved then cooled to 0° C. andDIAD (116 mg, 0.574 mmol, 113 μL) was added dropwise with mixing. Themixture was warmed to 35° C. and stirred for 18 hours. Flashchromatography (ISCO system, silica (12 g), 0-80% ethyl acetate inhexane) provided 2-1. (81.1 mg, 155 mol, 56% yield).

Step 2. To a solution of 2-1 (81.1 mg, 155 μmol) in DCM (1.5 mL) wasadded HCl in 1,4-dioxane (4 M, 1.5 mL). The mixture was stirred ambienttemperature for 1 hour, concentrated under reduced pressure, and driedunder high vacuum to provide 2-2.

Step 3. To a solution of 2-2 (65.6 mg, 155 μmol) in toluene (3.1 mL) wasadded triemethylaluminum in THF (2 M, 465 μL). The mixture was heated to100° C. and stirred for 1 hour. The mixture was cooled to roomtemperature and quenched with 2.0N aqueous HCl (4 mL), diluted withwater (10 mL) and extracted with ethyl acetate (3×10 mL). Combinedorganic layers was washed with brine and dried over sodium sulfate andconcentrated under reduced pressure. Flash chromatography (ISCO system,silica (12 g), 0-10% methanol in dichloromethane) provided 2. (29.0 mg,77 mol, 49% yield).

Compound 3 was prepared according to General Method I.

General Method J.

Preparation of(7S)-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile(4)

Step 1. A-3 was converted to 4-1 following step 1 in general method H.

Step 2. 4-1 was converted to 4-2 following step 2 in general method H.

Step 3. To a degassed mixture of 4-2 (8.0 mg, 17.9 μmol), Zn(CN)₂ (10.5mg, 8i9.2 μmol), Zn (0.12 mg, 1.8 μmol) and dppf (3.96 mg, 7.14 μmol) inDMA (1.12 mL) was added Pd₂(dba)₃ (3.3 mg, 3.6 μmol). The mixture washeated to 130° C. for 3 hours. The reaction was cooled and water (3 mL)added followed by extracted with dichloromethane (3×3 mL) Combinedextracts were dried with Na₂SO₄ then concentrated under reducedpressure. Flash chromatography (ISCO system, silica (12 g), 0-5%methanol in dichloromethane) followed by reverse phase purification ISCOsystem, C18 (50 g, gold), 0-100% acetonitrile in water w/0.035% TFA)provided 4 (6.7 g, 16.7 mol, 95% yield).

Compound 5 through 9 were prepared according to General Method I and Jstarting with A-4 through A-8 respectively.

General Method K.

Preparation of tert-butyl2-amino-5-[(4-methylbenzene-1-sulfonyl)oxy]pyrazolo[1,5-a]pyrimidine-3-carboxylate(A-10-5)

Step 1. To a solution of A-10-1 (1.58 kg, 15.0 mol, 1.60 L, 1.0 eq.) andtriethylamine (82.2 g, 812 mmol, 113 mL, 0.054 eq.) in ethanol (4.1 L)was added A-3-1A (3.80 kg, 26.30 mol, 2.64 L, 1.75 eq.) slowly. Themixture was stirred at 0-25° C. for 3 hours. The mixture wasconcentrated to give crude product. The residue was triturated withmixture solvent (2.0 L×3, PE:EA=5:1, V/V), Then the mixture was filteredand the filter cake was concentrated to give A-10-2 (2.78 kg, 9.74 mol,65% yield) as a white solid. ¹H NMR (400 MHz, CHLOROFORM-d) 6=10.20 (brs, 1H), 6.80 (br s, 1H), 1.55 (s, 9H).

Step 2. To a solution of A-10-2 (2.26 kg, 7.91 mol, 1.0 eq.) in dimethylformamide (4.1 L) was added NH₂NH₂H₂O (1.91 kg, 19.0 mol, 1.85 L, 50% inwater, 2.40 eq.). The mixture was stirred at 100° C. for 6 hours. Themixture was cooled to room temperature and concentrated to give compoundA-10-3 (2.7 kg, crude) as a black brown oil. ¹H NMR (400 MHz, DMSO-d₆)δ=5.29 (br s, 2H), 3.39 (br s, 2H), 1.47 (s, 9H).

Step 3. To a solution of A-10-3 (1020 g, 3.70 mol, 1.0 eq.) and A-3-3A(480 g, 3.43 mol, 0.926 eq.) in t-BuOH (6.0 L) was added sodium ethoxide(1.02 kg, 15 mol, 4.05 eq. fresh Prepared). The mixture was stirred at90° C. for 6 hours. The mixture was dissolved in ice-water (6.0 L) andquenched by acetic acid (2 M, 2.5 L) to neutralize PH=6 and extractedwith dichloromethane (3.5 L×5). The organic layer was washed by brine(5.0 L×3) and dried over anhydrous sodium sulfate. The solvent wasconcentrated to give crude product and the crude product was trituratedby solvent (3 L, PE:EA=1:1). The suspension was filtered and the filtercake was concentrated to give A-10-4 (704 g, 2.68 mol, 72.31% yield, 96%purity) as a yellow solid. ¹H NMR (400 MHz, CHLOROFORM-d) 6=7.83 (d,J=8.0 Hz, 1H), 5.95 (d, J=8.0 Hz, 1H), 4.94 (br s, 2H), 1.62 (s, 9H).

Step 4. To a solution of A-10-4 (987 g, 3.79 mol, 1.0 eq.) indichloromethane (6.0 L) was added triethylamine (1.51 kg, 14.9 mol, 2.08L, 3.93 eq.) and paratoluensulfonyl chloride (750 g, 3.93 mol, 1.04eq.). The mixture was stirred at 0° C.−25° C. for 5 hours. The mixturewas cooled to room temperature and concentrated to give crude product.The crude product was dissolved in dichloromethane (5.0 L) and washed bywater (4.0 L×3). The organic layer was concentrated to give productcompound A-10-5 (1.14 kg, 2.80 mol, 73.79% yield, 95.9% purity) as apink solid. ¹H NMR (400 MHz, CHLOROFORM-d) 6=8.32 (d, J=7.2 Hz, 1H),8.17 (d, J=8.4 Hz, 2H), 7.35 (d, J=8.0 Hz, 2H), 6.50 (d, J=7.2 Hz, 1H),5.38 (s, 2H), 2.45 (s, 3H), 1.65 (s, 9H).

General Method L

Preparation of(7S)-3-amino-14-(²H₅)ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile(10)

Step 1. A solution of A-2-4 (1.0 g, 4.57 mmol), 10-1A (1.14 g, 4.79mmol) and K₂CO₃ (1.89 g, 13.7 mmol) in DMF (15 mL) was stirred for 3hours at 25° C. The reaction mixture was diluted with DCM (100 mL) andwater (75 mL) and adjusted till acidic with 20% citric acid solution andstirred vigorously for 10 min. The organic layer was removed and theaqueous layer extracted with DCM (2×25 mL). The combined extracts werewashed with brine (50 mL), dried over anhydrous sodium sulfate, filteredand concentrated to dryness. Flash chromatography (ISCO system, silica(80 g), 10-40% ethyl acetate in hexane) provided 10-1 (1.70 g, 99%yield).

Step 2. A solution of 10-1 (4.09 g, 10.9 mmol) and 10-2A (1.8 g, 35.9mmol) in dry methanol (54 mL) was stirred for 1 hour at 50° C. Reactionwas cooled to room temperature and NaBH₄ (822 mg, 21.7 mmol) was added.The reaction mixture was stirred for 14 hours then quenched with water(75 mL). The mixture was extracted with DCM (3×75 mL). The combinedextracts were washed with brine (50 mL), dried over anhydrous sodiumsulfate, filtered and concentrated. Flash chromatography (ISCO system,silica (40 g), 10-80% ethyl acetate in hexane) provided 10-2 (4.05 g,90% yield).

Step 3. To a mixture of A-10-5 (2.8 g, 6.92 mmol), 10-2 (2.98 g, 7.27mmol) and molecular sieve (3 g) in n-butanol (10.0 mL) was added DIEA(4.47 g, 34.6 mmol). The mixture was heated to 90° C. and stirred for 26hours. The reaction was cooled and diluted with DCM (100 mL) thenfiltered through celite. The filtrate was washed with 1 M Na₂CO₃solution (50 mL) then brine (50 mL) and dried over anhydrous sodiumsulfate, filtered and concentrated. Flash chromatography (ISCO system,silica (120 g), 0-60% ethyl acetate in dichloromethane) provided 10-3(4.07 g, 91% yield).

Step 4. To a degassed solution of 10-3 (4.07 g, 6.33 mmol) in DMF (12.6mL) was added CuCN (850 mg, 9.5 mmol). The mixture was heated to 110° C.and stirred for 39 hours. The reaction was cooled and diluted with DCM(15 mL) then 6 M NH₄OH solution (50 mL) was added. The mixture wasstirred vigorously for 15 min then extracted with DCM (4×35 mL) andcombined extracts were again mixed vigorously with a 6 M NH₄OH solution(50 mL) for 30 min extracted with DCM (3×50 mL) and repeated treatmentwith NH₄OH solution 2 more time. Combined extracts were dried with brine(50 mL) then Na₂SO₄ and concentrated under reduced pressure. Flashchromatography (ISCO system, silica (120 g), 20-60% ethyl acetate indichloromethane) followed by reverse phase purification (ISCO system,C18 (50 g, gold), 0-100% acetonitrile in water w/0.035% TFA, 6injections) provided 10-4 (2.82 g, 75% yield).

Step 5. To a solution of 10-4 (2.82 mg, 4.80 mmol) in DCM (25 mL) wasadded HCl in 1,4-dioxane (4 M, 20 mL, 80 mmol). The mixture was stirredambient temperature for 16 hours, concentrated under reduced pressure,and dried under high vacuum. The crude material was dissolved in in DMF(10 mL) and DCM (60 mL) and Hünig's base (1.56 g, 120 mmol, 21 mL) thenFDPP (2.02 g, 5.27 mmol) was added in one portion. The reaction wasstirred for 87 hours then quenched with 2 M Na₂CO₃ solution (100 mL).Mixture was stirred for 5 min then extracted with DCM (3×150 mL).Combined extracts were washed with 2 M Na₂CO₃ solution (100 mL), brine(100 mL) and dried with Na₂SO₄ and concentrated under reduced pressure.Flash chromatography (ISCO system, silica (120 g), 1.25-6.25% methanolin dichloromethane) provided 10 (1.59 g, 79% yield).

General Method M

Preparation of(7R)-3-amino-14-ethyl-11-fluoro-7-methyl-4-oxo-4,5,6,7,13,14-hexahydro-1,15-ethenopyrazolo[4,3-f][1,4,8,10]benzoxatriazacyclotridecine-12-carbonitrile(11)

Step 1. To a solution of A-6-1 (438.42 g, 1.70 mol, 1.0 eq.) and A-10-5(690 g, 1.70 mol, 1.0 eq.) in n-BuOH (6.0 L) was addeddiisopropylethylamine (742 g, 5.74 mol, 1.0 L, 3.38 eq.) and 4A MS (200g). The mixture was stirred at 90° C. for 8 hours. TLC (PE:EA=1:1)showed the compound 7 was consumed and two new spots were found. Themixture was filtered at 50° C. and the filtrate was quenched by water(8.0 L) and extracted with ethyl acetate (4.0 L×3). The organic layerwashed by brine (4.0 L×3), dried over anhydrous sodium sulfate andconcentrated to give crude product. The filter cake was stirred inn-BuOH (2.0 L) at 90° C. for 1 hour, then filtered at 50° C., repeatedthis work up for three times until no desired product remained whichmonitored by TLC, then the filtrate was concentrated to give crudeproduct. All the residues were triturated with mixture solvent [500mL×3; ethyl actate:petroleum ether=1:2 (v/v)] and the mother liquid waspurified by column chromatography (SiO₂, Petroleum ether/Ethylacetate=10/1 to 0:1) to give 11-1 (570 g, 1.10 mol, 65.1% yield, 93.4%purity) as a yellow solid. ¹H NMR (400 MHz, CHLOROFORM-d) 6=10.31 (s,1H), 8.02 (d, J=7.6 Hz, 1H), 7.02 (dd, J=8.0, 8.8 Hz, 1H), 6.82 (dd,J=4.8, 8.8 Hz, 1H), 6.14 (d, J=7.6 Hz, 1H), 5.29 (s, 2H), 5.17 (br s,2H), 3.54 (q, J=7.2 Hz, 2H), 1.60 (s, 9H), 1.26 (t, J=7.2 Hz, 3H).

Step 2. To a solution of 11-1 (8.00 g, 16.7 mmol, 1.00 eq.) in dimethylformamide (25.0 mL) was added cuprous cyanide (2.24 g, 24.9 mmol, 5.46mL, 1.50 eq.). The mixture was stirred at 130° C. for 10 hours. Thereaction mixture was added ammonium hydroxide (10.0 mL) and diluted withwater (300 mL). Then the mixture was extracted with ethyl acetate (300mL×3). The combined organic layers were washed with saturated ammoniumchloride (100 mL×5), dried over saturated sodium sulfate, filtered andconcentrated under reduced pressure to give a residue, The residue waspurified by prep-HPLC (column: Phenomenex Gemini C18 250*50 mm*10 um;mobile phase: [water (0.05% ammonia hydroxide v/v)-ACN]; B %: 40%-60%,45MIN; 70% min) to give 11-2 (2.00 g, 4.54 mmol, 27.2% yield, 96.7%purity) as a brown solid. ¹H NMR (400 MHz, DMSO-d₆) δ=10.66 (br s, 1H),8.38 (d, J=7.6 Hz, 1H), 7.31 (t, J=9.2 Hz, 1H), 7.16 (dd, J=4.8, 8.8 Hz,1H), 6.52 (d, J=7.6 Hz, 1H), 5.96 (s, 2H), 4.94 (br s, 2H), 3.50 (br d,J=6.8 Hz, 2H), 1.46 (s, 9H), 1.10 (t, J=6.8 Hz, 3H).

Step 3. To a solution of 11-2 (2.05 g, 4.81 mmol, 1.00 eq.) in dimethylformamide (20.0 mL) was added potassium carbonate (1.66 g, 12.0 mmol,2.50 eq.) and 11-2A (1.71 g, 7.21 mmol, 1.50 eq.). The mixture wasstirred at 30° C. for 6 hours. The reaction mixture was diluted withwater (100 mL) and extracted with ethyl acetate (100 mL×3). The combinedorganic layers were washed with brine (100 mL×3), dried over anhydroussodium sulfate, filtered and concentrated under reduced pressure to givea residue, The residue was purified by prep-HPLC (column: PhenomenexGemini C18 250*50 mm*10 um; mobile phase: [water (0.05% ammoniahydroxide v/v)-ACN]; B %: 50%-80%, 25MIN80% min) to give 11-3 (1.80 g,3.05 mmol, 63.4% yield, 98.9% purity) as a light yellow solid. ¹H NMR(400 MHz, DMSO-d₆) δ=8.36 (d, J=8.0 Hz, 1H), 7.52-7.40 (m, 2H), 6.96 (brs, 1H), 6.52 (br d, J=7.6 Hz, 1H), 5.93 (s, 2H), 5.12-4.91 (m, 2H),4.58-4.44 (m, 1H), 3.44 (br s, 2H), 3.21-3.09 (m, 1H), 3.08-2.95 (m,1H), 1.44 (s, 9H), 1.34 (s, 9H), 1.11 (d, J=6.4 Hz, 3H), 1.07 (t, J=6.8Hz, 3H).

Step 4. 11-3 was converted to 11 following step 5 in General Method L.

¹H NMR (300 MHz, DMSO-d₆) δ Cpd Structure MS m/z ppm 1

419.1 8.84 (t, J = 4.68 Hz, 1 H) 8.35 (d, J = 7.70 Hz, 1 H) 7.18-7.30(m, 1 H) 7.07-7.18 (m, 1 H) 6.48 (d, J = 7.89 Hz, 1 H) 5.81 (s, 2 H)5.55 (dd, J = 15.04, 1.83 Hz, 1 H) 4.80-4.92 (m, 1 H) 3.97-4.13 (m, 2 H)3.67- 3.83 (m, 2 H) 3.22-3.29 (m, 1 H) 1.44 (d, J = 6.05 Hz, 3 H) 1.18(t, J = 6.97 Hz, 3 H) 2

377.2 9.05-8.95 (m, 1H), 8.78 (d, J = 7.9 Hz, 1H), 8.09 (s, 1H), 7.49(dd, J = 2.6, 7.0 Hz, 1H), 7.40-7.29 (m, 2H), 6.95 (d, J = 8.0 Hz, 1H),5.51 (d, J = 15.1 Hz, 1H), 5.02-4.86 (m, 1H), 4.39-4.13 (m, 2H),3.90-3.74 (m, 2H), 3.32-3.21 (m, 1H), 1.48 (d, J = 6.2 Hz, 3H),1.23-1.17 (m, 3H) 3

363.2 8.88 (dd, J = 3.3, 5.8 Hz, 1H), 8.78 (d, J = 7.9 Hz, 1H), 8.09 (s,1H), 7.51 (dd, J = 3.0, 6.6 Hz, 1H), 7.41-7.33 (m, 2H), 6.95 (d, J = 8.0Hz, 1H), 5.54 (d, J = 15.1 Hz, 1H), 4.72-4.59 (m, 1H), 4.51 (ddd, J =5.3, 9.2, 11.6 Hz, 1H), 4.35-4.13 (m, 2H), 3.83 (dd, J = 7.2, 15.3 Hz,1H), 3.73-3.49 (m, 2H), 1.20 (t, J = 7.0 Hz, 3H) 4

395.2 8.93 (t, J = 4.9 Hz, 1H), 8.80 (d, J = 7.9 Hz, 1H), 8.10 (s, 1H),7.55 (dd, J = 4.7, 9.4 Hz, 1H), 7.32 (t, J = 8.9 Hz, 1H), 6.96 (d, J =8.0 Hz, 1H), 5.55-5.43 (m, 1H), 4.99-4.85 (m, 1H), 4.34 (d, J = 15.3 Hz,1H), 4.28-4.15 (m, 1H), 3.38-3.28 (m, 1H), 1.47 (d, J = 6.1 Hz, 3H),1.20 (t, J = 7.0 Hz, 3H) 5

410.2 8.69 (t, J = 5.00 Hz, 1 H) 8.41 (d, J = 7.61 Hz, 1 H) 7.53 (dd, J= 9.49, 4.72 Hz, 1 H) 7.26-7.37 (m, 1 H) 6.60 (d, J = 7.79 Hz, 1 H)5.86-6.14 (m, 2 H) 5.45 (br d, J = 15.41 Hz, 1 H) 4.85-4.97 (m, 1 H)4.26 (d, J = 15.50 Hz, 1 H) 4.02-4.17 (m, 1 H) 3.64-3.80 (m, 2 H)3.23-3.35 (m, 1 H) 1.46 (d, J = 6.24 Hz, 3 H) 1.17 (t, J = 6.92 Hz, 3 H)6

392.2 8.76 (t, J = 4.86 Hz, 1 H) 8.39 (d, J = 7.61 Hz, 1 H) 7.48 (dd, J= 6.97, 2.75 Hz, 1 H) 7.29-7.37 (m, 2 H) 6.58 (d, J = 7.79 Hz, 1 H) 5.48(d, J = 15.04 Hz, 1 H) 4.86-5.00 (m, 1 H) 4.21 (d, J = 15.13 Hz, 1 H)4.03- 4.17 (m, 1 H) 3.68-3.83 (m, 2 H) 3.29 (dt, J = 13.04, 5.03 Hz, 1H) 1.47 (d, J = 6.14 Hz, 3 H) 1.17 (t, J = 6.97 Hz, 3 H) 7

436.2 8.73 (t, J = 5.09 Hz, 1 H) 8.39 (d, J = 7.70 Hz, 1 H) 7.53 (dd, J= 9.49, 4.81 Hz, 1 H) 7.31 (t, J = 8.89 Hz, 1 H) 6.71 (d, J = 7.70 Hz, 1H) 5.45- 5.57 (m, 1 H) 4.83-4.94 (m, 1 H) 4.38 (br d, J = 15.13 Hz, 2 H)4.22 (br dd, J = 15.36, 5.64 Hz, 3 H) 3.73 (dt, J = 13.53, 4.93 Hz, 1 H)3.23- 3.37 (m, 2 H) 1.47 (d, J = 6.14 Hz, 3 H) 1.02-1.17 (m, 1 H)0.43-0.57 (m, 3 H) 0.21-0.31 (m, 1 H) 8

424.2 8.39-8.51 (m, 2 H) 7.55 (dd, J = 9.54, 4.68 Hz, 1 H) 7.28 (t, J =8.80 Hz, 1 H) 6.73 (d, J = 7.70 Hz, 1 H) 5.17 (br d, J = 15.77 Hz, 1 H)4.95- 5.04 (m, 2 H) 4.48 (dt, J = 13.02, 6.33 Hz, 2 H) 4.33 (br d, J =15.68 Hz, 1 H) 3.59-3.71 (m, 1 H) 3.30 (ddd, J = 13.89, 6.05, 3.07 Hz, 1H) 1.58 (d, J = 6.51 Hz, 3 H) 1.48 (d, J = 6.24 Hz, 3 H) 1.16 (d, J =6.33 Hz, 3 H) 9

382.2 9.32-9.43 (m, 1 H) 8.65 (br t, J = 5.41 Hz, 1 H) 8.25 (d, J = 7.43Hz, 1 H) 7.48 (dd, J = 9.49, 4.81 Hz, 1 H) 7.28-7.40 (m, 1 H) 6.29 (d, J= 7.34 Hz, 1 H) 5.15 (br dd, J = 14.86, 2.66 Hz, 1 H) 4.66-4.79 (m, 1 H)4.10- 4.22 (m, 1 H) 3.77-3.87 (m, 1 H) 3.15-3.25 (m, 1 H) 1.44 (d, J =6.05 Hz, 3 H) 10

415.3 8.69 (t, J = 4.91 Hz, 1 H) 8.41 (d, J = 7.70 Hz, 1 H) 7.54 (dd, J= 9.49, 4.81 Hz, 1 H) 7.23-7.39 (m, 1 H) 6.60 (d, J = 7.70 Hz, 1 H) 5.85(s, 2 H) 5.44 (dd, J = 15.13, 1.38 Hz, 1 H) 4.82-5.00 (m, 1 H) 4.26 (d,J = 15.22 Hz, 1 H) 3.70 (dt, J = 13.64, 4.37 Hz, 1 H) 3.22-3.32 (m, 1 H)1.46 (d, J = 6.24 Hz, 3 H) 11

410.1 8.69 (br t, J = 4.72 Hz, 1 H) 8.41 (d, J = 7.61 Hz, 1 H) 7.53 (dd,J = 9.40, 4.72 Hz, 1 H) 7.31 (t, J = 8.89 Hz, 1 H) 6.60 (d, J = 7.70 Hz,1 H) 5.85 (s, 2 H) 5.44 (br d, J = 15.31 Hz, 1 H) 4.83-5.00 (m, 1 H)4.26 (br d, J = 15.31 Hz, 1 H) 4.00-4.18 (m, 1 H) 3.61-3.81 (m, 2 H)3.23-3.31 (m, 1 H) 1.46 (d, J = 6.24 Hz, 3 H) 1.17 (t, J = 6.88 Hz, 3 H)

Large Scale Preparation of Compound 5

Charged the reactor with A-10-5 (1.0 eq), A-6-1 (1.1 eq), DIPEA (3.0 eq)and n-butanol (10 vol). The resulting mixture was heated at 90-95° C.for 10 h. The reaction progress was monitored by HPLC, 2% of A-10-5revealed that the reaction is complete. After a passing IPC test thereaction mixture was cooled to 0-5° C. The reaction mixture was stirredfor 2 hours. The solids were filtered and washed with cold MTBE (2×0.5volume). The solids were dried under vacuum oven at 40-50° C. toconstant weight with yield 2950 g (75%) and 99.9% HPLC purity.

Charged the reactor with 11-1 (1.0 eq), DMA (5 vol) and CuCN (2.5 eq).The resulting mixture was heated at 90-100° C. for 92 hours. Thereaction progress was monitored by HPLC, NMT 2% of 11-1 revealed thatthe reaction is complete. After a passing IPC test. Transferred thereaction mixture into 2^(nd) reactor containing DCM (46 L, 15 vol),Celite (3073 g) at 35-40° C. The reaction mixture was stirred for 30 minat 20-30° C. Filtered the reaction mixture through one inch celite bedand washed with DCM (2×5 vol). Filtrate was charged with Celite (3073g), charcoal (1000 g) and Buffer (H₂O/NH₄Cl/NH₄OH, 9.4/4.0/3.8; 31 L, 10v) to filtrate. The reaction mixture was stirred for 2 hours at 20-30°C. Filtered the reaction mixture through one inch celite bed and washedwith DCM (2×5 vol). Separated the layers and the organic layer waswashed with Buffer (2×31 L, 2×10 v) and water (2×10 vol). Concentratedthe organic to minimum volume and co-evaporated with MTBE (2×5 vol). Thereaction mixture was cooled to 15-30° C. and stirred for 4 hours. Thesolids were filtered and washed with cold Methanol (2×1 volume). Thesolids were dried under vacuum oven at 40-50° C. to constant weight withyield 2310 g (82%) and 99% HPLC purity.

Charged the reactor with 11-2 (1.0 eq), 10-1A (1.15 eq), acetonitrile (5vol) and DBU (2.5 eq). The resulting mixture was stirred at 20-30° C.for 2 hours. The reaction progress was monitored by HPLC, 1% of 11-2revealed that the reaction is complete. After a passing IPC test. Thereactor was charged ethyl acetate (10 vol) and 25 wt % citric acidsolution (10 vol). The reaction mixture was stirred overnight andseparated the layers and back extracted the aqueous layer with Ethylacetate (10 volumes). Combined organic layer was concentrated to minimumvolume and co-evaporated with DCM (2×5 vol). Concentrated to drynessobtained 1630 g (92%) and 99% HPLC purity.

Charged the reactor with 5-A (1.0 eq), DCM (10 vol) and 4M HCl inDioxane (10 eq). The resulting mixture was stirred at 20-30° C. for 2hours. The reaction progress was monitored by HPLC, 1% of 5-A revealedthat the reaction is complete. After a passing IPC test. The solids werefiltered, washed with MTBE (2×5 vol) and dried the solids in filter withvacuum under nitrogen to obtained yield with 1450 g (Assume 100% 1300 g)and 97% purity.

Charged the reactor with 5-B (1.0 eq), DIPEA (5.0 eq) and DCM (20 vol)and DMF (1 vol). The resulting mixture was stirred at room temperaturefor (15-30° C.) for 15-30 min. Charged reactor with FDPP (1.3 eq) in oneportion. The resulting mixture was stirred at room temperature for(15-30° C.) overnight. The reaction progress was monitored by HPLC, 1%of 5-B revealed that the reaction is complete. After a passing IPC test.The reactor was charged 1M Na₂CO₃ solution (10 volumes). The reactionmixture was stirred for 30 min. and separated the layers. The organiclayer was washed with 1M Na₂CO₃ solution (10 volumes) and water (2×10vol) and brine (50 volumes). The organic layer was concentrated theorganic to minimum volume and co-evaporated with methanol (2×5 vol).Organic layer was dried with MgSO₄ and charcoal and Filtered the organiclayer though GF paper and concentrated the organic to minimum volume andco-evaporated with ethanol (2×5 vol). Concentrated to dryness and addedEtOH (2 L) and stir at room temperature for 1 hour. The solids werefiltered and washed with cold Methanol (2×1 volume). The solids weredried under vacuum oven at 40-50° C. to constant weight with yield 850 g(86%).

Charged the reactor with crude 5 (1.0 eq) and water (12 vol). Theresulting mixture was stirred at room temperature for 3 days. The solidswere filtered and washed with water (2×1 volume). The solids were driedunder vacuum oven at 40-50° C. to constant weight with yield 723 g (86%)and 98.7% HPLC purity.

Biologic Assays

In-Vitro Assays

Materials and Methods

Biochemical Kinase Assay Method

The biochemical kinase assay was performed at Reaction BiologyCorporation (www.reactionbiology.com, Malvern, Pa.) following theprocedures described in the reference (Anastassiadis T, et al NatBiotechnol. 2011, 29, 1039). Specific kinase/substrate pairs along withrequired cofactors were prepared in reaction buffer; 20 mM Hepes pH 7.5,10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na3VO4, 2mM DTT, 1% DMSO (for specific details of individual kinase reactioncomponents see Supplementary Table 2). Compounds were delivered into thereaction, followed 20 minutes later by addition of a mixture of ATP(Sigma, St. Louis Mo.) and ³³P ATP (Perkin Elmer, Waltham Mass.) to afinal concentration of 10 μM. Reactions were carried out at roomtemperature for 120 min, followed by spotting of the reactions onto P81ion exchange filter paper (Whatman Inc., Piscataway, N.J.). Unboundphosphate was removed by extensive washing of filters in 0.75%phosphoric acid. After subtraction of background derived from controlreactions containing inactive enzyme, kinase activity data was expressedas the percent remaining kinase activity in test samples compared tovehicle (dimethyl sulfoxide) reactions. IC₅₀ values and curve fits wereobtained using Prism (GraphPad Software).

Cell Lines and Cell Culture:

Human gastric cancer cell line SNU-5, lung cancer cell lines HCC827,H1975, mouse myelogenous leukemia cell line M-NFS-60 were obtained fromATCC. Cell lines Ba/F3, MKN-45 were purchased from DSMZ. SNU-216 cellline was purchased from KCLB.

Cloning and Ba/F3 Stable Cell Line Creation

The TEL-CSF-1R cDNA was synthesized at GenScript and cloned intopCDH-CMV-MCS-EF1-Puro plasmid (System Biosciences, Inc). Ba/F3 TEL-CSF1Rwas generated by transducing Ba/F3 cells with lentivirus containingTEL-CSF1R cDNA clone. Stable cell lines were selected by puromycintreatment, followed by IL-3 withdrawal. Briefly, 1×10⁶ Ba/F3 cells weretransduced with lentivirus supernatant in the presence of 8 μg/mLprotamine sulfate. The transduced cells were subsequently selected with1 μg/mL puromycin in the presence of IL3-containing medium RPMI1640,plus 10% FBS. After 10-12 days of selection, the surviving cells werefurther selected for IL3 independent growth.

Cell Proliferation Assays:

Two thousand cells per well were seeded in 384 well white plate for 24hrs, and then treated with compounds for 72 hours (37° C., 5% CO₂). Cellproliferation was measured using CellTiter-Glo luciferase-based ATPdetection assay (Promega) following the manufacturer's protocol. IC₅₀determinations were performed using GraphPad Prism software (GraphPad,Inc., San Diego, Calif.).

Immunoblotting for Cellular Kinase Phosphorylation Assays

Gastric carcinoma cell lines MKN-45, SNU-5 (both with METoverexpression), HCC827 cells (harboring endogenous EGFR mutationdelE1746_A750), NCI-H1975 cells (harboring endogenous EGFR doublemutations L858R/T790M) or SNU216 cells were cultured in RPMI 1640medium, supplemented with 10% fetal bovine serum and 100 U/mL ofpenicillin/streptomycin. Half a million cells per well were seeded in 24well plate for 24 hrs, and then treated with compounds for 4 hours.Cells were collected after treatment and lysed in RIPA buffer (50 mMTris, pH 7.4, 150 mM NaCl, 1% NP-40, 0.5% Deoxycholate, 0.1% SDS)supplemented with 10 mM EDTA, 1× Halt protease and phosphataseinhibitors (Thermo Scientific). Protein lysates (approximately 20 μg)was resolved on 4-12% Bolt Bis-Tris precasted gels with MES runningbuffer (Life Technologies), transferred to nitrocellulose membranesusing Trans-Blot Turbo Transfer System (Bio-Rad) and detected withantibodies targeting phosphorylated MET (Y1234/Y1235) (Cell SignalingTechnology), MET (Y1349), MET (Y1003), total MET (Cell SignalingTechnology), phosphorylated EGFR (Y1068) and total EGFR(Cell SignalingTechnology), phosphorylated STAT3 and STATS, total STAT3 and STATS (CellSignaling Technology), phosphorylated AKT (Cell Signaling Technology),total AKT (Cell Signaling Technology), phosphorylated ERK (CellSignaling Technology), total ERK (Cell Signaling Technology),phosphorylated PLCγ2 and total PLCγ2 (Cell Signaling Technology),phosphorylated SRC Y416 (Cell Signaling Technology), total SRC (CellSignaling Technology), phosphorylated paxillin Y118 (Cell SignalingTechnology), total paxillin (Cell Signaling Technology), PARP, actin(Cell Signaling Technology). Antibodies were typically incubatedovernight at 4° C. with gentle shake, followed by washes and incubationwith the appropriate HRP-conjugated secondary antibodies. Membranes wereincubated with chemiluminescent substrate for 5 min at room temperature(SuperSignal West Femto, Thermo Scientific). The chemiluminescent imageswere acquired with a C-DiGit Imaging System (LI-COR Biosciences). Therelative density of the chemiluminescent bands were quantified via ImageStudio Digits from LICOR. The half inhibitory concentration (IC₅₀) valueis calculated using non-linear regression analysis through GraphPadPrism software (GraphPad, Inc., San Diego, Calif.).

The Scratch Wound Healing Assays

MKN-45 or HCC827 cells were seeded in 24-well plate. After 12-24 hours,confluent cell monolayers were gently scraped with a sterile pipette tipto form a scratch. The plates were washed with fresh medium, and thecells were incubated with medium alone or medium containing variousconcentration of compounds. After 36-48 hours, the plates were examinedand recorded by an EVOS FL microscopy (Life Technology) to monitorresealing of the cell monolayer.

In-Vivo Methods

Cell Lines

MKN-45 and Ba/F3 ETV6-CSF1R cells were cultured using standardtechniques in RPMI-1640 medium (Corning, Inc) with 10% fetal bovineserum (Thermo Fisher Scientific, Inc) at 37° C. in a humidifiedatmosphere with 5% CO₂. For implantation, cells were harvested andpelleted by centrifugation at 250 g for 2 minutes. Cells were washedonce and resuspended in serum-free medium supplemented with 50% matrigel(v/v).

Subcutaneous Xenograft Models in Immune Compromised Mice

Female athymic nude mice (5-8 weeks of age) were obtained from CharlesRiver Laboratory and were housed in Innovive IVC disposable cages onHEPA filtered ventilated racks with ad libitum access to rodent chow andwater. Five million cells in 100 μL serum-free medium supplemented with50% matrigel (Corning, Inc) were implanted subcutaneously in the rightflank region of the mouse. Tumor size and body weight were measured ondesignated days. Tumor size was measured with an electronic caliper andtumor volume was calculated as the product of length*width²*0.5. Micewere randomized by tumor size into treatment groups when tumor volumereached about 200 mm³ and Compound 5 was administered orally (BID) atdetermined doses.

Tumor Processing and Immunoblotting for In Vivo Pharmacodynamic Studies

Mice bearing xenograft tumors were humanely euthanized and tumors wereresected and snap frozen in liquid nitrogen and stored at −80° C. Frozentumor samples were processed at 4° C. in 1× Cell Lysis Buffer (CellSignaling Technologies) to extract proteins. SDS loading samples wereprepared by adding one volume of 4×LDS Sample Buffer (Life Technologies,Inc) to three volumes of protein lysate. Tumor SDS protein samples wereprocessed by SDS-PAGE and immunoblotted with rabbit anti-phosphorylatedMET, mouse anti-MET and mouse anti-actin antibodies (Cell SignalingTechnologies). The signals from immunoblot were detected by C-DiGit BlotScanner from LI-COR and the signal intensity were quantified using theImage Studio Digit software (LI-COR).

Subcutaneous Patient-Derived Xenograft Model in Immune Compromised Mice

Female BALB/c nude mice (6-7 weeks) were obtained from Beijing AnikeeperBiotech Co. Ltd (Beijing, China). Primary human tumor xenograft modelLU2503 tumors were grown in stock mice. Tumor fragments (2-3 mm indiameter) were harvested from stock mice and inoculated into the rightfront back of each mouse for tumor development. 16 mice were enrolled inthe study. All animals were randomly allocated to the 2 different studygroups. Tumor size and body weight were measured on designated days.Tumor size was measured using a caliper and tumor volume was calculatedas the product of length*width²*0.5. Mice were randomized by tumor sizeinto treatment groups when tumor volume reached about 200 mm³ andCompound 5 was administered orally (BID) at 15 mg/kg.

Subcutaneous MC38 Syngeneic Model in C57BL/6J Mice

C57BL/6J female mice (6 weeks) were purchased from the JacksonLaboratory, and maintained in accordance with the guidelines for thecare and use of laboratory animals. Half million MC38 cancer cells in100 μL serum-free medium were implanted subcutaneously in the rightflank region of the mouse. Tumor size and body weight were measured ondesignated days. Tumor size was measured with an electronic caliper andtumor volume was calculated as the product of length*width²*0.5. Micewere randomized by tumor size into treatment groups when tumor volumereached about 70-90 mm³. Vehicle control, Compound 5, PD-1 antibody orCompound 5 plus PD-1 antibody were administered orally (BID) atdetermined doses.

MC38 Syngeneic Model PD Biomarker Studies

MC38 tumors were collected on day 7 and day 11. The collected tumorswere dissociated using MiltenyiGentleMax. FACS analysis of tumors wereperformed for the tumor associated immune cells including tumorassociated macrophages (TAM) and TAM subtypes (M1 and M2), myeloidderived suppression cells (MDSC), cytotoxic T lymphocytes (CTL, i.e.CD8+ T cells), CD4+ T cells, and regulatory T cells (Treg).

Data and Results:

Enzymatic Kinase Activities

The enzymatic kinase inhibition activities at 10 μM ATP concentrationwere determined at Reaction Biology. The results of IC₅₀ were summarizedin Table 1.

TABLE 1 Enzymatic Enzymatic Enzymatic Kinase Kinase Kinase c-FMS (CSF1R)SRC IC₅₀ MET IC₅₀ IC₅₀ Compd # (nM) (nM) (nM) 1 ND* 20.4 ND 2 3.00 36.5ND 3 97.1 110.0 ND 4 0.49 1.85 0.42 5 0.12 0.14 0.76 6 13.6 9.7 ND 70.70 2.2 ND 8 0.83 3.8 ND *ND = not determined

Anti-Cell Proliferation Activities

The anti-cell proliferation activities against MET and CSF1R driven celllines were conducted with MKN-45, SNU-5, Ba/F3 TEL-CSF1R cells, andM-NFS-60 respectively. The results of IC₅₀ were summarized in Table 2and Table 3.

TABLE 2 MET Cell MET Cell CSF1R Cell Proliferation ProliferationProliferation Ba/F3 MKN-45 IC₅₀ SNU-5 IC₅₀ TEL-CSF1R IC₅₀ Compd # (nM)(nM) (nM) 1 193 173.9 180.3 2 129 135.9 281.4 3 471 674.1 1740 4 12 5.898.1 5 0.2 0.17 19.3 6 58.3 36.8 187.7 7 17.7 1.0 108.1 8 1.0 1.0 39.0 9297 10 0.2 11 251

TABLE 3 M-NFS-60 CSF-1 (ng/mL) (IC₅₀s nM) 0 0.3 1 3 10 30 100Pexidartinib <0.1 2 146.4 212.5 379.7 594.7 702.3 (PLX-3397) Compound 50.3 3 11.6 78.2 84.1 180.8 174.5

Compound 5 Inhibited the Phosphorylation of MET and Downstream Signaling

The pharmacodynamic inhibiting activity of Compound 5 on MET and thecorresponding downstream signaling in MET-driven cells was evaluated,and the results were shown in FIGS. 1 and 2. Compound 5 caused thesuppression of MET autophosphorylation as well as the downstream STAT3,ERK and AKT phosphorylation at IC₅₀s of around 1-3 nM in SNU-5 andMKN-45 cell lines (FIGS. 1 &2).

Compound 5 Synergized with AZD9291 in HCC827 Cells

The lung cancer cell line HCC827 has endogenous EGFR exon 19 deletionwith MET overexpression. The EGFR inhibitor AZD9291 showed an IC₅₀ of 5nM, however, with a maximum inhibition of Emax 47% in cell proliferationassay. The selective MET inhibitor capmatinib is not active in HCC827cell proliferation assay. The combination of AZD9291 with capmatinibshowed a similar effect as AZD9291 alone with an IC₅₀ of 5 nM and Emax48%. The MET/SRC dual inhibitor Compound 5 showed an IC₅₀ of 3000 nM inHCC827 cell proliferation assay. A strong synergistic activity wasobserved in the combination of AZD9291 with Compound 5 with an IC₅₀ of 2nM and Emax 71% in HCC827 cell proliferation assay. The results weresummarized in FIG. 3. Compound 5 synergized with AZD9291 for apoptosisin HCC827 cell line as shown in FIG. 4.

Evaluation of the Migration Inhibition of Compound 5

Compound 5 inhibited the migration of MKN-45 or HCC827 cells after 36-48hours treatment in the wound healing assays, whereas, the selective METinhibitor capmatinib only inhibited the migration of MKN-45 cells andhas minimum effect on HCC827 cells. The results were presented in FIGS.5 & 6.

In-Vivo Studies

Antitumor Efficacy of Compound 5 in Xenograft Tumor Models

The antitumor efficacy of Compound 5 was evaluated in several tumorxenograft models representing cancer populations in which dysregulationof MET is implicated.

MKN-45 Gastric Adenocarcinoma Model

The Met gene amplification in MKN-45 cells underlies the molecularmechanism for tumor growth. Athymic nude mice bearing MKN-45 tumors (atthe average tumor size of 210 mm³) were dosed with Compound 5 orally BIDfor twelve days (FIG. 7). The control group of mice were given vehicleonly. Tumor volume (TMV) was measured by caliper on the indicated daysand is shown at mean±sem in FIG. 7. The mean TMVs are significantlylower in the treated groups compared to that of the control group(p<0.05) as determined by two-way repeated ANOVA followed by post hocanalysis. Tumor growth inhibition (TGI) was calculated as100%*1{−[(TMV_(Treated Last Day of Treatment)−TMV_(Treated First Day of Treatment))/(TMV_(Control on Last Day of Treatment)−TMV_(Control on First Day of Treatment))]}whenTMV_(Treated Last Day of Treatment)≥TMV_(Treated First Day of Treatment).In the case ofTMV_(Treated Last Dayof Treatment)<TMV_(Treated First Day of Treatment),tumor regression (REG) was calculated as100%*(1−TMV_(Treated Last Day of Treatment)/TMV_(Treated First Day of Treatment)).In this study, Compound 5 demonstrated the ability to inhibit tumorgrowth at 47% at the dose of 3 mg/kg, BID. When dosed at 10 mg/kg, BIDand 30 mg/kg, BID, treatment of Compound 5 resulted in 6% and 44% tumorregression, respectively. Tumor size was reduced in 5 out 10 micetreated with Compound 5 at 10 mg/kg, BID and in 9 out of 10 mice treatedwith Compound 5 at 30 mg/kg. Body weight of the mice were measured onthe designated days of the mice as shown in FIG. 8.

Inhibition of MET Activity in MKN-45 Tumors Following OralAdministration of Compound 5

To evaluate the effect of Compound 5 on the inhibition of METphosphorylation, MKN-45 tumors were harvested at either 0.5 hour afteran oral dose of Compound 5 at 10 mg/kg. The level of MET phosphorylationwas determined by immunoblotting combined with signal quantification bythe Image Studio Digit Software. Compound 5 inhibited METphosphorylation to 16% and 13% of the control level at Tyr-1234 andTyr-1349, respectively (FIG. 9). In another experiment, tumors wereharvested after repeated dose administration at 4 hours and 12 hoursafter last dose of Compound 5. The level of MET phosphorylation atTyr-1234 was determined by ELISA. Compound 5 inhibited METphosphorylation to 0.2% and 4.0% of control level at 4 hours and 12hours after last dose of 10 mg/kg Compound 5 treatment; Compound 5inhibited MET phosphorylation to 12.7% and 33.1% of control level at 4hours and 12 hours after last dose of 10 mg/kg Compound 5 treatment(FIG. 10).

LU2503 Patient Derived Xenograft (PDX) NS CLC Model

The LU2503 is a PDX model derived from a NSCLC patient and harboringgene amplification and exon 14 skipping mutation of the Met gene.Treating mice bearing LU2503 tumors with Compound 5 at 15 mg/kg, BID for13 days resulted in a 85% tumor regression, whereas the tumors grew from189 mm³ to 2032 mm³ in the vehicle treated group (FIG. 11). No bodyweight loss was observed after 21 days of BID treatment with Compound 5at 15 mg/kg (FIG. 12).

Inhition of the Growth of Ba/F3 ETV6-CSF1R Tumors

In the Ba/F3 ETV6-CSF1R xenograft tumor model, the growth of tumor ispresumably dependent on the extopic CSF1R activity. SCID/Beige micebearing Ba/F3 ETV6-CSF1R tumors with average tumor size of 180 mm³) weredosed with Compound 5 orally BID for 10 days (FIG. 13). The controlgroup of mice were given vehicle only. Tumor volume (TMV) was measuredby caliper on the indicated days and is shown at mean±sem in FIG. 12.The mean TMVs are significantly lower in the treated groups compared tothat of the control group (p<0.05) as determined by two-way repeatedANOVA followed by post hoc analysis. Compound 5 demonstrated the abilityto inhibit tumor growth at 44% and 67% at the dose of 5 mg/kg, BID and15 mg/kg, BID, respectively. Body weight of the mice were measured onthe designated days of the mice as shown in FIG. 14.

The PD Marker Evaluation of Compound 5 in the Subcutaneous MC38Syngeneic Mouse Tumor Model

Anti-tumor effects of Compound 5 on MC38 syngeneic tumors was analyzedby tumor volume. The average tumor volume of vehicle control group (G1)on day 7 was 696.3±299.7, while Compound 5 treated group (G2) was473.5±170.4 mm³. On day 11, the average tumor volume of G1 and G2 were1142.6±290.0 and 610.4±151.8 mm³, respectively. On day 11, tumor volumeshowed statistically significant difference between treatment groupswith p<0.006, while the difference was not statistically significant onDay 7. Percent tumor volume change is shown in FIG. 15. No body weightloss and overt abnormality was observed in mice treated with Compound 5at 15 mg/kg BID for 7 or 11 days, as showed in FIG. 16.

FACS analysis of tumors were performed on day 7 and day 11 for the tumorassociated immune cells including tumor associated macrophages (TAM) andTAM subtypes (M1 and M2), myeloid derived suppression cells (MDSC),cytotoxic T lymphocytes (CTL, i.e. CD8+ T cells), CD4+ T cells, andregulatory T cells (Treg). Data are shown in FIGS. 17 and 18. On day 7,there were no statistically significant changes in the populations ofTAM, M1, M2, MDSC, CTL, CD4+ T cells, or Treg in tumor associatedleukocytes (CD45+ populations) between the control and Compound 5treated groups, although there is a trend for a reduction in TAM cellsin Compound 5 treated mice. However, on day 11, a statisticallysignificant decrease of TAM in the total tumor leukocyte population wasobserved in the Compound 5 treated group compared to the control group,with a concurrent increase in the MDSC populations. Further analysis thesubpopulation of TAM revealed an increase in M1 TAM and a decrease in M2TAM in the total tumor leukocyte population in tumors in Compound 5treated group compared to the control group. At the same time, a trendof increase of CTL cells in the total tumor leukocyte population and astatistically significant increase of CTL in the CD3+ lymphocytepopulation were observed in the Compound 5 treated group compared to thecontrol group, with no statistically significant change found in theCD4+ T cells or Treg cells.

In Vivo Combination Efficacy Study of Compound 5 with PD-1 Antibody inMC38 Syngeneic Model

Anti-tumor effects of Compound 5 combined with PD-1 antibody on MC38syngeneic tumors was analyzed by tumor volume. The average tumor volumeof vehicle control group (G1) on day 20 was 1938.58±729.41, Compound 5treated group (G2) was 1220.03±521.39 mm3, PD-1 antibody treatment groupwas 821.24±767.16, and Compound 5 plus PD-1 antibody treatment was515.63±350.47. On day 20, an anti-tumor synergy was observed comparedthe combination group to the groups of Compound 5 or PD-1 antibodytreated alone. No body weight loss and overt abnormality was observed inmice treated Compound 5 and/or PD-1 antibody. Data are shown in FIGS. 19and 20.

1. A compound to of the Formula I

or a pharmaceutically acceptable salt thereof, wherein X¹ and X² areindependently —CR⁶R⁷—, S, S(O), S(O)₂, O or N(R⁸); IV is H, deuterium,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl, C₃-C₁₀aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein each hydrogen atom in C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl and C₆-C₁₀ aryl isindependently optionally substituted by deuterium, halogen, —OH, —CN,—OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl,—N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆alkyl)₂, —N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl,—N(C₁-C₆ alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆alkyl), —N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆alkyl), —NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆ alkyl)₂, —P(O)(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or 3-to 7-membered heterocycloalkyl; each R² and R³ is independently H,deuterium, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl,C₆-C₁₀ aryl, —C(O)OR⁸ or —C(O)NR⁸R⁹; wherein each hydrogen atom in C₁-C₆alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl and C₆-C₁₀ aryl isindependently optionally substituted by deuterium, halogen, —OH, —CN,—OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, —NHC(O)C₁-C₆alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂, —NHC(O)NHC₁-C₆ alkyl,—N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆ alkyl, —NHC(O)N(C₁-C₆alkyl)₂, —N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂, —NHC(O)OC₁-C₆ alkyl,—N(C₁-C₆ alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆ alkyl), —NHS(O)₂(C₁-C₆alkyl), —N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂(C₁-C₆alkyl), —NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆ alkyl)S(O)NH₂, —N(C₁-C₆alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl), —NHS(O)₂NH(C₁-C₆ alkyl),—NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H,—C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂,—SC₁-C₆ alkyl, —S(O)C₁-C₆ alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆alkyl), —S(O)₂NH(C₁-C₆ alkyl), —S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆alkyl)₂, —P(C₁-C₆ alkyl)₂, —P(O)(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or 3-to 7-membered heterocycloalkyl; or R² and R³ taken together with thecarbon atoms to which they are attached optionally form a C₅-C₇cycloalkyl or a 5- to 7-membered heterocycloalkyl; or R² and R⁴ takentogether with the atoms to which they are attached optionally form a 5-to 7-membered heterocycloalkyl; R⁴ is H, C₁-C₆ alkyl or 3- to 7-memberedheterocycloalkyl, wherein each hydrogen atom in C₁-C₆ alkyl or 3- to7-membered heterocycloalkyl is independently optionally substituted byhalogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)₂, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl),—C(O)N(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or monocyclic 5- to 7-memberedheterocycloalkyl; R⁵ is —NR⁶R⁷; each R⁶, R⁷ and R⁸ are eachindependently selected from the group consisting of H, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₂-C₆ alkynyl, and C₃-C₆ cycloalkyl; wherein eachhydrogen atom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, and C₃-C₆cycloalkyl is independently optionally substituted by deuterium, fluoro,chloro, bromo, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)₂, C₃-C₇ cycloalkyl, 3- to 7-membered heterocycloalkyl, C₆-C₁₀aryl, 5- to 7-membered heteroaryl, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl), or —C(O)N(C₁-C₆ alkyl)₂; R⁹ is fluoro, chloro,bromo, —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆alkyl) and —C(O)N(C₁-C₆ alkyl)₂; R¹⁰ is H, fluoro, chloro or bromo; andn is 1 or 2; with the proviso that when R⁵ is H, R⁹ is selected from thegroup consisting of —CN, —CF₃, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl) and —C(O)N(C₁-C₆ alkyl)₂. 2.-5. (canceled)
 6. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R⁶ and R⁷ are H.
 7. (canceled)
 8. The compound of claim 6, or apharmaceutically acceptable salt thereof, wherein R⁹ is —CN.
 9. Thecompound of claim 8, or a pharmaceutically acceptable salt thereof,wherein R¹⁰ is fluoro.
 10. The compound of claim 9, or apharmaceutically acceptable salt thereof, wherein X¹ is N(R⁸).
 11. Thecompound of claim 10, or a pharmaceutically acceptable salt thereof,wherein R⁸ is C₁-C₆ alkyl, wherein each hydrogen atom is independentlyoptionally substituted by fluoro, chloro, bromo, —OH, —CN, —OC₁-C₆alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)₂, C₃-C₇ cycloalkyl, 3- to7-membered heterocycloalkyl, C₆-C₁₀ aryl, 5- to 7-membered heteroaryl,—CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂, —C(O)NH(C₁-C₆ alkyl), or—C(O)N(C₁-C₆ alkyl)₂.
 12. The compound of claim 10, or apharmaceutically acceptable salt thereof, wherein R⁸ is ethyl, propyl,iso-propyl, or methylcyclopropyl.
 13. The compound of claim 11, or apharmaceutically acceptable salt thereof, wherein X² is O.
 14. Thecompound of claim 12, or a pharmaceutically acceptable salt thereof,wherein R² is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆cycloalkyl, C₆-C₁₀ aryl, —C(O)OR⁷ or —C(O)NR⁷R⁸; wherein each hydrogenatom in C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₃-C₆ cycloalkyl andC₆-C₁₀ aryl is independently optionally substituted by deuterium,halogen, —OH, —CN, —OC₁-C₆ alkyl, —NH₂, —NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)₂, —NHC(O)C₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)C₁-C₆ alkyl, —NHC(O)NH₂,—NHC(O)NHC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)NH₂, —N(C₁-C₆ alkyl)C(O)NHC₁-C₆alkyl, —NHC(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆ alkyl)C(O)N(C₁-C₆ alkyl)₂,—NHC(O)OC₁-C₆ alkyl, —N(C₁-C₆ alkyl)C(O)OC₁-C₆ alkyl, —NHS(O)(C₁-C₆alkyl), —NHS(O)₂(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)(C₁-C₆ alkyl),—N(C₁-C₆alkyl)S(O)₂(C₁-C₆ alkyl), —NHS(O)NH₂, —NHS(O)₂NH₂, —N(C₁-C₆alkyl)S(O)NH₂, —N(C₁-C₆ alkyl)S(O)₂NH₂, —NHS(O)NH(C₁-C₆ alkyl),—NHS(O)₂NH(C₁-C₆ alkyl), —NHS(O)N(C₁-C₆ alkyl)₂, —NHS(O)₂N(C₁-C₆alkyl)₂, —N(C₁-C₆ alkyl)S(O)NH(C₁-C₆ alkyl), —N(C₁-C₆alkyl)S(O)₂NH(C₁-C₆ alkyl), —N(C₁-C₆ alkyl)S(O)N(C₁-C₆ alkyl)₂, —N(C₁-C₆alkyl)S(O)₂N(C₁-C₆ alkyl)₂, —CO₂H, —C(O)OC₁-C₆ alkyl, —C(O)NH₂,—C(O)NH(C₁-C₆ alkyl), —C(O)N(C₁-C₆ alkyl)₂, —SC₁-C₆ alkyl, —S(O)C₁-C₆alkyl, —S(O)₂C₁-C₆ alkyl, —S(O)NH(C₁-C₆ alkyl), —S(O)₂NH(C₁-C₆ alkyl),—S(O)N(C₁-C₆ alkyl)₂, —S(O)₂N(C₁-C₆ alkyl)₂, —P(C₁-C₆ alkyl)₂,—P(O)(C₁-C₆ alkyl)₂, C₃-C₆ cycloalkyl, or 3- to 7-memberedheterocycloalkyl, and R³ is H.
 15. The compound of claim 14, or apharmaceutically acceptable salt thereof, wherein R² is C₁-C₆ alkyl. 16.The compound of claim 15, or a pharmaceutically acceptable salt thereof,wherein R² is methyl. 17.-18. (canceled)
 19. The compound of claim 1,selected from the group consisting of

or a pharmaceutically acceptable salt thereof.
 20. A pharmaceuticalcomposition comprising a compound of claim 1, or a pharmaceuticallyacceptable salt thereof, and at least one or more of a pharmaceuticallyacceptable diluent, carrier or excipient.
 21. A method of treatingcancer in a patient comprising, a administering a therapeuticallyeffective amount of a compound that inhibits SRC and MET, and/or CSF1R.22. The method of claim 22, wherein the compound that inhibits SRC andMET, and/or CSF1R is of the formula of claim
 1. 23. The method of claim22, wherein the cancer is gastric cancer, colon cancer, renal cancer,liver cancer, lung cancer, glioblastoma, or head & neck cancer.
 24. Themethod of claim 22, further comprising b. administering atherapeutically effective amount of at least one additional anti-canceragent.
 25. The method of claim 24, wherein the at least one additionalanti-cancer agent is an EGFR inhibitor, or a pharmaceutically acceptablesalt thereof.
 26. The method of claim 24, wherein the additionalanti-cancer agent is an antibody of EGFR.
 27. The method of claim 26,wherein the antibody of EGFR is cetuximab, necitumumab or panitumumab.28. The method of claim 24, wherein the additional anti-cancer agent isa small molecule inhibitor of EGFR.
 29. The method of claim 28, whereinthe small molecule inhibitor of EGFR is afatinib, brigatinib,canertinib, dacomitinib, erlotinib, gefitinib, HKI 357, lapatinib,osimertinib, naquotinib, nazartinib, neratinib, olmutinib, pe itinib,PF-06747775, rociletinib, vandetanib, or a pharmaceutically acceptablesalt thereof. 30.-80. (canceled)